钩吻老茎与嫩茎生物碱成分的差异分析
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摘要
利用UPLC-Q-TOF/MS~E结合主成分分析(PCA)及正交偏最小二乘判别分析(OPLS-DA),研究了钩吻老茎和嫩茎生物碱类成分的差异,找出了17个主要的差异性成分,并明确了差异性生物碱在老茎及嫩茎中的分布,其中极性较大的生物碱在钩吻嫩茎中含量较高,而在老茎中,极性较小的生物碱含量较高。同时利用HPLC-UV定量分析了钩吻中3个主要的生物碱。结果表明老茎中的钩吻素子和钩吻素己含量高于嫩茎,而嫩茎中钩吻素甲含量较高。该研究系统分析了钩吻老茎及嫩茎生物碱类成分的组成及差异,同时对其3个主要的生物碱成分进行了定量分析,为钩吻药材安全有效的应用奠定了基础。
This study aimed to comprehensively assess the difference of alkaloid components between old stems and tender stems of Gelsemium elegans by using ultra high-performance liquid chromatography coupled with photo-diode array and quadrupole time-of-flight mass spectrometry( UPLC-Q-TOF/MS~E) and high-performance liquid chromatography coupled with UV detector( HPLC-UV). Firstly,the different components in old stems and tender stems were analyzed by UHPLC-Q-TOF/MSEcombined with principal component analysis( PCA) and orthogonal partial least squares discriminant analysis( OPLS-DA),respectively. As a result,17 major different components were found. At the same time,the distribution of these alkaloids in old stems and tender stems was determined,and the alkaloids with higher polarity are relatively higher in the tender stems,while the old stems are in the opposite case. In addition,three main components in the G. elegans were quantified by HPLC-UV. The results showed that the contents of koumine and humantenmine in old stems were higher than those in tender stems,and the content of gelsemine in tender stems was relatively high. This study systematically evaluated the differences of alkaloids between the old stems and tender stems of G. elegans,and quantified the main three alkaloids. It laid the foundation of the safe and effective application of G. elegans.
This study aimed to comprehensively assess the difference of alkaloid components between old stems and tender stems of Gelsemium elegans by using ultra high-performance liquid chromatography coupled with photo-diode array and quadrupole time-of-flight mass spectrometry( UPLC-Q-TOF/MS~E) and high-performance liquid chromatography coupled with UV detector( HPLC-UV). Firstly,the different components in old stems and tender stems were analyzed by UHPLC-Q-TOF/MSEcombined with principal component analysis( PCA) and orthogonal partial least squares discriminant analysis( OPLS-DA),respectively. As a result,17 major different components were found. At the same time,the distribution of these alkaloids in old stems and tender stems was determined,and the alkaloids with higher polarity are relatively higher in the tender stems,while the old stems are in the opposite case. In addition,three main components in the G. elegans were quantified by HPLC-UV. The results showed that the contents of koumine and humantenmine in old stems were higher than those in tender stems,and the content of gelsemine in tender stems was relatively high. This study systematically evaluated the differences of alkaloids between the old stems and tender stems of G. elegans,and quantified the main three alkaloids. It laid the foundation of the safe and effective application of G. elegans.
引文
[1]中国科学院中国植物志委员会.中国植物志.第61卷[M].北京:科学出版社,1992:251.
[2] Xu Y K,Liao S G,Na Z,et al. Gelsemium alkaloids,immunosuppressive agents from Gelsemium elegans[J]. Fitoterapia,2012,83(6):1120.
[3]黄静,许盈,俞昌喜,等.钩吻生物碱化合物抗肿瘤效应及其诱导凋亡作用[J].海峡药学,2014,26(10):19.
[4]黄静,苏燕评,俞昌喜,等.钩吻生物碱化合物体外抗消化系统肿瘤的活性[J].海峡药学,2010,22(3):197.
[5]谭建权,邱成之,郑林忠.钩吻碱的镇痛作用和无依赖性[J].中药药理与临床,1988(1):24.
[6]俞昌喜,许盈,苏燕评,等.钩吻生物碱钩吻素子的提纯及其抗神经病理性疼痛的作用[J].中国药理学与毒理学杂志,2012,26(3):443.
[7]刘浩,俞昌喜.钩吻的研究进展[J].福建医科大学学报,2008,42(5):469.
[8] Jin G L,Su Y P,Liu M,et al. Medicinal plants of the genus Gelsemium(Gelsemiaceae,Gentianales)--a review of their phytochemistry,pharmacology, toxicology and traditional use[J].J Ethnopharmacol,2014,152(1):33.
[9]冯时茵,栗建明,侯惠婵,等.钩吻生物碱成分的薄层色谱研究[J].中药新药与临床药理,2013,24(1):77.
[10]陈卫琳,杨樱,吴水生.闽产钩吻根茎叶中钩吻碱甲和钩吻素子及葫蔓藤碱甲的含量测定[J].福建中医药大学学报,2011,21(5):48.
[11] Li Y,Zeng R J,Lu Q,et al. Ultrasound/microwave-assisted extraction and comparative analysis of bioactive/toxic indole alkaloids in different medicinal parts of Gelsemium elegans Benth by ultra-high performance liquid chromatography with MS/MS[J].J Sep Sci,2014,37(3):308.
[12]栗建明,冯时茵,何华红,等.单一内标多控法同步测定钩吻中钩吻碱甲和钩吻素子的含量[J].中药材,2013,36(10):1626.
[13] Guo L X,Li R,Liu K,et al. Structural characterization and discrimination of Chinese medicinal materials with multiple botanical origins based on metabolite profiling and chemometrics analysis:Clematidis Radix et Rhizoma as a case study[J].J Chromatogr A,2015,1425:129.
[14]郑伟,王芳旭,孙欣光,等.硫磺熏蒸知母的化学标志物分析[J].中国中药杂志,2017,42(18):3530.
[15] Li H F,Guan X Y,Ye M,et al. Qualitative and quantitative analyses of Epimedium wushanense by high-performance liquid chromatography coupled with diode array detection and electrospray ionization tandem mass spectrometry[J]. J Sep Sci,2011,34(12):1437.
[16] Liu Z,Zhu Z,Zhang H,et al. Qualitative and quantitative analysis of Fructus Corni using ultrasound assisted microwave extraction and high performance liquid chromatography coupled with diode array UV detection and time-of-flight mass spectrometry[J].J Pharm Biomed Anal,2011,55(3):557.
[17] Zhang J Y,Gong N,Huang J L,et al. Gelsemine,a principal alkaloid from Gelsemium sempervirens Ait.,exhibits potent and specific antinociception in chronic pain by acting at spinalα3 glycine receptors[J]. Pain,2013,154(11):2452.
[18] Chen C J,Zhong Z F,Xin Z M,et al. Koumine exhibits anxiolytic properties without inducing adverse neurological effects on functional observation battery,open-field and Vogel conflict tests in rodents[J]. J Nat Med,2017,71(2):397.
[19] Ming L,Jie S,Hao L,et al. Gelsenicine from Gelsemium elegans attenuates neuropathic and inflammatory pain in mice[J]. Biol Pharm Bull,2011,34(12):1877.
[20] Liu Y C,Lin L,Cheng P,et al. Fingerprint analysis of Gelsemium elegans by HPLC followed by the targeted identification of chemical constituents using HPLC coupled with quadrupole-timeof-flight mass spectrometry[J]. Fitoterapia,2017,121:94.
[21] Liu Y C,Xiao S,Yang K,et al. Comprehensive identification and structural characterization of target components from Gelsemium elegans by high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry based on accurate mass databases combined with MS/MS spectra[J]. J Mass Spectrom,2017,52(6):378.
[22]张琳,赵庆春,李艳.钩吻生物碱成分的研究进展[J].沈阳药科大学学报,2007(8):515.
[23]张桢,刘光明,何红平.钩吻吲哚生物碱类化合物研究进展[J].大理学院学报,2008(6):10.
[2] Xu Y K,Liao S G,Na Z,et al. Gelsemium alkaloids,immunosuppressive agents from Gelsemium elegans[J]. Fitoterapia,2012,83(6):1120.
[3]黄静,许盈,俞昌喜,等.钩吻生物碱化合物抗肿瘤效应及其诱导凋亡作用[J].海峡药学,2014,26(10):19.
[4]黄静,苏燕评,俞昌喜,等.钩吻生物碱化合物体外抗消化系统肿瘤的活性[J].海峡药学,2010,22(3):197.
[5]谭建权,邱成之,郑林忠.钩吻碱的镇痛作用和无依赖性[J].中药药理与临床,1988(1):24.
[6]俞昌喜,许盈,苏燕评,等.钩吻生物碱钩吻素子的提纯及其抗神经病理性疼痛的作用[J].中国药理学与毒理学杂志,2012,26(3):443.
[7]刘浩,俞昌喜.钩吻的研究进展[J].福建医科大学学报,2008,42(5):469.
[8] Jin G L,Su Y P,Liu M,et al. Medicinal plants of the genus Gelsemium(Gelsemiaceae,Gentianales)--a review of their phytochemistry,pharmacology, toxicology and traditional use[J].J Ethnopharmacol,2014,152(1):33.
[9]冯时茵,栗建明,侯惠婵,等.钩吻生物碱成分的薄层色谱研究[J].中药新药与临床药理,2013,24(1):77.
[10]陈卫琳,杨樱,吴水生.闽产钩吻根茎叶中钩吻碱甲和钩吻素子及葫蔓藤碱甲的含量测定[J].福建中医药大学学报,2011,21(5):48.
[11] Li Y,Zeng R J,Lu Q,et al. Ultrasound/microwave-assisted extraction and comparative analysis of bioactive/toxic indole alkaloids in different medicinal parts of Gelsemium elegans Benth by ultra-high performance liquid chromatography with MS/MS[J].J Sep Sci,2014,37(3):308.
[12]栗建明,冯时茵,何华红,等.单一内标多控法同步测定钩吻中钩吻碱甲和钩吻素子的含量[J].中药材,2013,36(10):1626.
[13] Guo L X,Li R,Liu K,et al. Structural characterization and discrimination of Chinese medicinal materials with multiple botanical origins based on metabolite profiling and chemometrics analysis:Clematidis Radix et Rhizoma as a case study[J].J Chromatogr A,2015,1425:129.
[14]郑伟,王芳旭,孙欣光,等.硫磺熏蒸知母的化学标志物分析[J].中国中药杂志,2017,42(18):3530.
[15] Li H F,Guan X Y,Ye M,et al. Qualitative and quantitative analyses of Epimedium wushanense by high-performance liquid chromatography coupled with diode array detection and electrospray ionization tandem mass spectrometry[J]. J Sep Sci,2011,34(12):1437.
[16] Liu Z,Zhu Z,Zhang H,et al. Qualitative and quantitative analysis of Fructus Corni using ultrasound assisted microwave extraction and high performance liquid chromatography coupled with diode array UV detection and time-of-flight mass spectrometry[J].J Pharm Biomed Anal,2011,55(3):557.
[17] Zhang J Y,Gong N,Huang J L,et al. Gelsemine,a principal alkaloid from Gelsemium sempervirens Ait.,exhibits potent and specific antinociception in chronic pain by acting at spinalα3 glycine receptors[J]. Pain,2013,154(11):2452.
[18] Chen C J,Zhong Z F,Xin Z M,et al. Koumine exhibits anxiolytic properties without inducing adverse neurological effects on functional observation battery,open-field and Vogel conflict tests in rodents[J]. J Nat Med,2017,71(2):397.
[19] Ming L,Jie S,Hao L,et al. Gelsenicine from Gelsemium elegans attenuates neuropathic and inflammatory pain in mice[J]. Biol Pharm Bull,2011,34(12):1877.
[20] Liu Y C,Lin L,Cheng P,et al. Fingerprint analysis of Gelsemium elegans by HPLC followed by the targeted identification of chemical constituents using HPLC coupled with quadrupole-timeof-flight mass spectrometry[J]. Fitoterapia,2017,121:94.
[21] Liu Y C,Xiao S,Yang K,et al. Comprehensive identification and structural characterization of target components from Gelsemium elegans by high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry based on accurate mass databases combined with MS/MS spectra[J]. J Mass Spectrom,2017,52(6):378.
[22]张琳,赵庆春,李艳.钩吻生物碱成分的研究进展[J].沈阳药科大学学报,2007(8):515.
[23]张桢,刘光明,何红平.钩吻吲哚生物碱类化合物研究进展[J].大理学院学报,2008(6):10.