UPLC-Q-TOF-MS/MS技术研究了哥王水提物中的化学成分
|
摘要
该研究应用超高效液相色谱-飞行时间质谱(UPLC-Q-TOF-MS/MS)技术对了哥王水提物中化学成分进行定性分析。采用AcclaimTMRSLC 120-C18色谱柱(2. 1 mm×100 mm,2. 2μm);以0. 2%甲酸铵缓冲盐溶液(A)-0. 2%甲酸铵缓冲盐甲醇溶液(B)为流动相进行梯度洗脱;柱温30℃;采用电喷雾离子源,正、负离子全扫描模式采集数据。通过二级高分辨质谱分析结合对照品数据及相关文献,从了哥王水提物中共鉴定出52个化合物,包括11个香豆素类、13个黄酮类、10个木脂素类、2个酰胺类、4个酚酸类、6个倍半萜类和6个其他类化合物,并首次从了哥王中发现indicanone同分异构体、β-hydroxypropiovanillone和epiprocurcumenol 3个化合物。此外,该研究对一些化合物的质谱裂解规律进行推断总结,并首次阐述愈创木烷型倍半萜类化合物的裂解规律,为研究类似化学成分的质谱裂解途径提供参考。该研究采用UPLC-Q-TOF-MS/MS技术,为鉴定中药复杂化学成分提供了简单快速的方法,为进一步研究了哥王化学成分奠定基础。
In this study,a method using ultra performance liquid chromatography tandem quadrupole time-of-flight mass spectrometry( UPLC-Q-TOF-MS/MS) was established to identify complicated chemical constituents of Wikstroemia indica. Chromatographic separation was performed on an AcclaimTMRSLC 120-C18 column( 2. 1 mm×100 mm,2. 2 μm) using gradient elution with 0. 2% ammonium formate buffer salt solution( A)-0. 2% ammonium formate buffer salt solution methanol( B) as mobile phase. The column temperature was maintained at 30 ℃. The analytes were determined by positive and negative ion modes with electro-spray ionization source. A total of 52 compounds( including eleven coumarins,thirteen flavonoids,ten lignans,two amides,four phenolic acids,six sesquiterpenes and six other compounds) were identified or tentatively characterized from the water extract of W. indica by comparing their retention times and MS spectra with those of authentic standards or literature datas. Three compounds were found for the first time from W.indica namely isomer of indicanone,β-hydroxypropiovanillone and epiprocurcumenol. Furthermore,the fragmentation rules of some compounds were speculated and summarized. In addition,the cleavage pathways of guaiane sesquiterpenes were described for the first time,which can provide reference for studying the fragmentation pathways of similar compounds. This study provides an easy way to identify chemical constituents of traditional Chinese medicine and a basis for the further study on chemical fundamentals of W. indica.
In this study,a method using ultra performance liquid chromatography tandem quadrupole time-of-flight mass spectrometry( UPLC-Q-TOF-MS/MS) was established to identify complicated chemical constituents of Wikstroemia indica. Chromatographic separation was performed on an AcclaimTMRSLC 120-C18 column( 2. 1 mm×100 mm,2. 2 μm) using gradient elution with 0. 2% ammonium formate buffer salt solution( A)-0. 2% ammonium formate buffer salt solution methanol( B) as mobile phase. The column temperature was maintained at 30 ℃. The analytes were determined by positive and negative ion modes with electro-spray ionization source. A total of 52 compounds( including eleven coumarins,thirteen flavonoids,ten lignans,two amides,four phenolic acids,six sesquiterpenes and six other compounds) were identified or tentatively characterized from the water extract of W. indica by comparing their retention times and MS spectra with those of authentic standards or literature datas. Three compounds were found for the first time from W.indica namely isomer of indicanone,β-hydroxypropiovanillone and epiprocurcumenol. Furthermore,the fragmentation rules of some compounds were speculated and summarized. In addition,the cleavage pathways of guaiane sesquiterpenes were described for the first time,which can provide reference for studying the fragmentation pathways of similar compounds. This study provides an easy way to identify chemical constituents of traditional Chinese medicine and a basis for the further study on chemical fundamentals of W. indica.
引文
[1]张婕斐,裘建社,徐新峰,等.了哥王片治疗急性扁桃体炎、急性咽炎、急性气管-支气管炎的临床观察[J].中国医院用药评价与分析,2014,14(3):248.
[2] Li Y M,Zhu L,Jiang J G,et al. Bioactive components and pharmacological action of Wikstroemia indica(L.)C. A. Mey. and its clinical application[J]. Curr Pharm Biotechnol,2009,10(8):743.
[3]李雨田,顾雪竹,张村.了哥王的化学成分和药理作用研究进展[J].中国实验方剂学杂志,2011,17(24):252.
[4]张鑫.了哥王的化学成分及镇痛抗炎活性研究[D].广州:广东药学院,2011.
[5]国光梅,汪冶,李玮,等.了哥王石油醚提取部位化学成分研究[J].科学技术与工程,2014,14(21):188.
[6]张立,刘慧琼,肖玥,等.了哥王叶子化学成分研究(I)[J].亚太传统医药,2012,8(8):41.
[7]邵萌,黄晓君,孙学刚,等.了哥王根茎中的酚性成分及其抗肿瘤活性研究[J].天然产物研究与开发,2014,26(6):851.
[8] Jiang Y B,Zhong M,Peng W. Qualitative analysis of plant-derived samples by liquid chromatography-electrospray ionizationquadrupole-time of flight-mass spectrometry[J]. Trop J Pharm Res,2015,14(5):925.
[9]彭耀文,申兰慧,王丽,等.液相色谱-四级杆-飞行时间质谱联用技术在药物分析中的应用[J].中南药学,2015(9):962.
[10]慕善学,孙丽霞,魏岚,等.HPLC同时测定了哥王中芦荟大黄素和大黄素甲醚的含量[J].中国实验方剂学杂志,2015,21(18):73.
[11]杨贤英,李金花,段宝忠,等.五倍子抑制流感病毒神经氨酸酶活性及活性部位的UPLC-Q-TOF-MS分析[J].中国实验方剂学杂志,2017,23(16):68.
[12]李晓庄,赵慧玲,黄文平,等.UPLC-Q-TOF-MS/MS快速鉴别北豆根中的生物碱类成分[J].中国实验方剂学杂志,2017,23(15):97.
[13]苏日娜,罗维早,魏荣锐,等.基于UPLC-ESI-Q-TOF-MS/MS技术的特色藏族药高原荨麻化学成分快速识别研究[J].中国中药杂志,2019,44(8):1607.
[14]尹震花,张娟娟,陈林,等.了哥王化学成分与生物活性研究进展[J].中草药,2018,49(8):1964.
[15]郑玉莹,庞文静,白杨,等.基于UFLC-Triple TOF-MS/MS技术的枳实传统饮片及破壁饮片化学成分系统分析[J].中南药学,2018,16(4):443.
[16]谢培山,杨景鹏.了哥王化学成分的研究-西瑞香素(Daphnoretin)的分离、鉴定[J].中草药通讯,1978(3):97.
[17] Cuyckens F,Claeys M. Mass spectrometry in the structural analysis of flavonoids[J]. J Mass Spectrom,2004,39(4):1.
[18] Fabre N,Rustan I,Hoffmann E D,et al. Determination of flavone,flavonol,and flavanone aglycones by negative ion liquid chromatography electrospray ion trap mass spectrometry[J]. J Am Soc Mass Spectrom,2001,12:707.
[19] Yang W Z,Ye M,Qiao X,et al. Collision-induced dissociation of40 flavonoid aglycones and differentiation of the common flavonoid subtypes using electrospray ionization ion-trap tandem mass spectrometry and quadrupole time-of-flight mass spectrometry[J]. Eur J Mass Spectrom,2012,18(6):493.
[20]王芦笛.基于液质联用技术的瑞香狼毒化学成分及药代动力学研究[D].石家庄:河北大学,2017.
[21]国光梅,李玮,汪冶,等.了哥王中木脂素成分及生物活性研究[J].山地农业生物学报,2012,31(5):457.
[22] Kato M,He Y M,Dibne D F,et al. New Guaian-type sesquiterpene from Wikstroemia indica[J]. Nat Prod Commun,2014,9(1):1.
[23] Wang L Y,Unehara T,Kitanaka S. Anti-inflammatory activity of new Guaiane type sesquiterpene from Wikstroemia indica[J].Chem Pharm Bull,2005,53(1):137.
[24] Jang M K,Lee H J,Kim J S,et al. A curcuminoid and two sesquiterpenoids from Curcuma zedoaria as inhibitors of nitric oxide synthesis in activated macrophages[J]. Arch Pharm Res,2004,27(12):1220.
[25] Tang J,Supinya T,Wang Z T,et al. Aurantiamide acetate from stems of Zanthoxylum dissitum Hemsley[J]. J Chin Pharm Sci,2003,12(4):231.
[26]耿立冬,张村,肖永庆.了哥王化学成分研究[J].中国中药杂志,2006,31(10):817.
[27]马尚智,秦惠玉,龙飞,等.没食子多酚类化合物的鉴定及其在UPLC-MS/MS中的裂解规律研究[J].中草药,2017,48(22):4632.
[28]孙国东,霍金海,程团,等.基于UPLC-Q-TOF/MS技术的胡桃楸叶化学成分分析[J].中药材,2017,40(5):1123.
[29]朱伟,赵赫,姚岚,等. UFLC-MS/MS法同时测定肺宁颗粒中6种化学成分[J].解放军药学学报,2017,33(4):356.
[30]殷会敏,黄文哲,肖伟.基于RRLC-Q-TOF-MS/MS技术快速分析芪葛颗粒的化学成分[J].中国中药杂志,2018,43(21):4240.
[31] Karonen M,Hmlinen M,Nieminen R,et al. Phenolic extractives from the bark of Pinus sylvestris L. and their effects on inflammatory mediators nitric oxide and prostaglandin E2[J]. J Agric Food Chem,2004,52(25):7532.
[2] Li Y M,Zhu L,Jiang J G,et al. Bioactive components and pharmacological action of Wikstroemia indica(L.)C. A. Mey. and its clinical application[J]. Curr Pharm Biotechnol,2009,10(8):743.
[3]李雨田,顾雪竹,张村.了哥王的化学成分和药理作用研究进展[J].中国实验方剂学杂志,2011,17(24):252.
[4]张鑫.了哥王的化学成分及镇痛抗炎活性研究[D].广州:广东药学院,2011.
[5]国光梅,汪冶,李玮,等.了哥王石油醚提取部位化学成分研究[J].科学技术与工程,2014,14(21):188.
[6]张立,刘慧琼,肖玥,等.了哥王叶子化学成分研究(I)[J].亚太传统医药,2012,8(8):41.
[7]邵萌,黄晓君,孙学刚,等.了哥王根茎中的酚性成分及其抗肿瘤活性研究[J].天然产物研究与开发,2014,26(6):851.
[8] Jiang Y B,Zhong M,Peng W. Qualitative analysis of plant-derived samples by liquid chromatography-electrospray ionizationquadrupole-time of flight-mass spectrometry[J]. Trop J Pharm Res,2015,14(5):925.
[9]彭耀文,申兰慧,王丽,等.液相色谱-四级杆-飞行时间质谱联用技术在药物分析中的应用[J].中南药学,2015(9):962.
[10]慕善学,孙丽霞,魏岚,等.HPLC同时测定了哥王中芦荟大黄素和大黄素甲醚的含量[J].中国实验方剂学杂志,2015,21(18):73.
[11]杨贤英,李金花,段宝忠,等.五倍子抑制流感病毒神经氨酸酶活性及活性部位的UPLC-Q-TOF-MS分析[J].中国实验方剂学杂志,2017,23(16):68.
[12]李晓庄,赵慧玲,黄文平,等.UPLC-Q-TOF-MS/MS快速鉴别北豆根中的生物碱类成分[J].中国实验方剂学杂志,2017,23(15):97.
[13]苏日娜,罗维早,魏荣锐,等.基于UPLC-ESI-Q-TOF-MS/MS技术的特色藏族药高原荨麻化学成分快速识别研究[J].中国中药杂志,2019,44(8):1607.
[14]尹震花,张娟娟,陈林,等.了哥王化学成分与生物活性研究进展[J].中草药,2018,49(8):1964.
[15]郑玉莹,庞文静,白杨,等.基于UFLC-Triple TOF-MS/MS技术的枳实传统饮片及破壁饮片化学成分系统分析[J].中南药学,2018,16(4):443.
[16]谢培山,杨景鹏.了哥王化学成分的研究-西瑞香素(Daphnoretin)的分离、鉴定[J].中草药通讯,1978(3):97.
[17] Cuyckens F,Claeys M. Mass spectrometry in the structural analysis of flavonoids[J]. J Mass Spectrom,2004,39(4):1.
[18] Fabre N,Rustan I,Hoffmann E D,et al. Determination of flavone,flavonol,and flavanone aglycones by negative ion liquid chromatography electrospray ion trap mass spectrometry[J]. J Am Soc Mass Spectrom,2001,12:707.
[19] Yang W Z,Ye M,Qiao X,et al. Collision-induced dissociation of40 flavonoid aglycones and differentiation of the common flavonoid subtypes using electrospray ionization ion-trap tandem mass spectrometry and quadrupole time-of-flight mass spectrometry[J]. Eur J Mass Spectrom,2012,18(6):493.
[20]王芦笛.基于液质联用技术的瑞香狼毒化学成分及药代动力学研究[D].石家庄:河北大学,2017.
[21]国光梅,李玮,汪冶,等.了哥王中木脂素成分及生物活性研究[J].山地农业生物学报,2012,31(5):457.
[22] Kato M,He Y M,Dibne D F,et al. New Guaian-type sesquiterpene from Wikstroemia indica[J]. Nat Prod Commun,2014,9(1):1.
[23] Wang L Y,Unehara T,Kitanaka S. Anti-inflammatory activity of new Guaiane type sesquiterpene from Wikstroemia indica[J].Chem Pharm Bull,2005,53(1):137.
[24] Jang M K,Lee H J,Kim J S,et al. A curcuminoid and two sesquiterpenoids from Curcuma zedoaria as inhibitors of nitric oxide synthesis in activated macrophages[J]. Arch Pharm Res,2004,27(12):1220.
[25] Tang J,Supinya T,Wang Z T,et al. Aurantiamide acetate from stems of Zanthoxylum dissitum Hemsley[J]. J Chin Pharm Sci,2003,12(4):231.
[26]耿立冬,张村,肖永庆.了哥王化学成分研究[J].中国中药杂志,2006,31(10):817.
[27]马尚智,秦惠玉,龙飞,等.没食子多酚类化合物的鉴定及其在UPLC-MS/MS中的裂解规律研究[J].中草药,2017,48(22):4632.
[28]孙国东,霍金海,程团,等.基于UPLC-Q-TOF/MS技术的胡桃楸叶化学成分分析[J].中药材,2017,40(5):1123.
[29]朱伟,赵赫,姚岚,等. UFLC-MS/MS法同时测定肺宁颗粒中6种化学成分[J].解放军药学学报,2017,33(4):356.
[30]殷会敏,黄文哲,肖伟.基于RRLC-Q-TOF-MS/MS技术快速分析芪葛颗粒的化学成分[J].中国中药杂志,2018,43(21):4240.
[31] Karonen M,Hmlinen M,Nieminen R,et al. Phenolic extractives from the bark of Pinus sylvestris L. and their effects on inflammatory mediators nitric oxide and prostaglandin E2[J]. J Agric Food Chem,2004,52(25):7532.