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
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摘要
采用超高效液相色谱-串联四极杆飞行时间质谱(UPLC-ESI-Q-TOF-MS/MS)技术对高原荨麻甲醇提取部分化合物进行了快速识别。采用Triple-TOF5600+高分辨质谱仪、超高效液相色谱、YMC-Triart C18色谱柱(2. 1 mm×100 mm,1. 9μm),乙腈-0. 4%甲酸水溶液为流动相梯度洗脱;质谱采用电喷雾(ESI)离子源,在负离子模式下采集数据,通过与对照品图谱中已知成分的保留时间、精确相对分子质量等比对,结合各成分的二级质谱裂解特征比对及文献报道的相同成分或同类成分的质谱碎片裂解规律分析,推测高原荨麻中成分的化学结构。从高原荨麻中初步鉴定出31个化合物,包括黄酮类化合物8个,酚酸类化合物14个,苯丙素类8个(香豆素类4个,木脂素类4个),萜类化合物1个,其中13个成分经对照品比对确认,以上化合物均为首次发现于高原荨麻中,为高原荨麻的药效物质基础等进一步深入研究奠定了基础。
This paper deals with the application of ultra-performance liquid chromatography tandem quadrupole time of flight mass spectrometry(UPLC-ESI-Q-TOF-MS/MS) method to rapidly determine and analyze the chemical constituents of methanol extract of Urtica hyperborea. We employed UPLC YMC-Triart C18(2. 1 mm×100 mm,1. 9 μm) column to UPLC analysis with acetonitrile-water(containing 0. 4% formic acid) in gradient as mobile phase. The flow rate was 0. 3 m L·min-1 gradient elution and column temperature was 30℃; the injection volume was 4 μL. ESI ion source was used to ensure the data collected in anegative ion mode. The chemical components of U. hyperborea were identified through retention time,exact relative molecular mass,cleavage fragments of MS/MS and reported data.The results indicated that a total of 31 compounds were identified,including 8 flavonoids,14 phenolic compounds,8 phenylpropanoids(4 coumarins and 4 lignans),and 1 steroidal compound,13 of which were confirmed by comparison. The UPLC-ESI-Q-TOF-MS/MS method could rapid identify the chemical components of U. hyperborea. The above compounds were discovered in U. hyperborea for the first time,which could provide theoretical foundation for further research on the basis of the pharmacodynamics of U. hyperborea.
This paper deals with the application of ultra-performance liquid chromatography tandem quadrupole time of flight mass spectrometry(UPLC-ESI-Q-TOF-MS/MS) method to rapidly determine and analyze the chemical constituents of methanol extract of Urtica hyperborea. We employed UPLC YMC-Triart C18(2. 1 mm×100 mm,1. 9 μm) column to UPLC analysis with acetonitrile-water(containing 0. 4% formic acid) in gradient as mobile phase. The flow rate was 0. 3 m L·min-1 gradient elution and column temperature was 30℃; the injection volume was 4 μL. ESI ion source was used to ensure the data collected in anegative ion mode. The chemical components of U. hyperborea were identified through retention time,exact relative molecular mass,cleavage fragments of MS/MS and reported data.The results indicated that a total of 31 compounds were identified,including 8 flavonoids,14 phenolic compounds,8 phenylpropanoids(4 coumarins and 4 lignans),and 1 steroidal compound,13 of which were confirmed by comparison. The UPLC-ESI-Q-TOF-MS/MS method could rapid identify the chemical components of U. hyperborea. The above compounds were discovered in U. hyperborea for the first time,which could provide theoretical foundation for further research on the basis of the pharmacodynamics of U. hyperborea.
引文
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[25]张靖,徐筱杰,徐文,等. HPLC-LTQ-Orbitrap-MSn快速鉴别党参药材中化学成分[J].中国实验方剂学杂志,2015,21(9):59.
[26] Fiorentino A,D'Abrosca B,Dellagreca M,et al. Chemical characterization of new oxylipins from Cestrum parqui, and their effects on seed germination and early seeding growth[J]. Chem Biodiv,2008,5(9):1780.
[27]李想.黄酮类化合物ESI-ITMSn质谱裂解规律的量子化学研究[D].佳木斯:佳木斯大学,2015.
[28]阿布拉江·克依木.黄酮苷类天然产物的质谱分析方法研究[D].北京:中国协和医科大学,2006.
[29]钟艳梅,冯毅凡,郭姣.基于UPLC-Q-TOF-MS技术的白术药材化学成分快速识别研究[J].质谱学报,2015,36(1):72.
[30]李蓉,秦坚民,余国莫.荨麻属药用植物研究概况及其开发价值[J].中国野生植物资源,2002,21(1):24.
[2]杨永昌.藏药志[M].西宁:青海人民出版社,1991:313.
[3]国家中医药管理局中华本草编委会.中华本草·藏药卷[M].上海:上海科学技术出版社,2002:238.
[4]罗达尚.新修晶珠本草[M].成都:四川科学技术出版社,2004:193.
[5]苏日娜,罗维早,朱继孝,等.荨麻属药用植物研究进展[J].中草药,2018,49(11):2722.
[6]袁昌齐.欧美植物药[M].南京:东南大学出版社,2004:18.
[7]伊智慧.咖啡酰奎宁酸类化合物ESI-IT MSn裂解的量子化学研究[D].佳木斯:佳木斯大学,2017.
[8] Chen S D,Lu C J,Zhao R Z. Identification and quantitative characterization of PSORI-CM01,a Chinese medicine formula for psoriasis therapy,by liquid chromatography coupled with an LTQ orbitrap mass spectrometer[J]. Molecules, 2015, 20:1594.
[9] Liu M H,Tong X,Wang J X,et al. Rapid separation and identification of multiple constituents in traditional Chinese medicine formula Shen-qi Fuzheng injection by ultra fast liquid chromatography combined with quadrupole time of flight mass spectrometry[J]. J Pharm Biomed Anal,2013,74:141.
[10]林颖,李康,陈华龙.曼胡颓子根的质量标准研究[J].中南药学,2015(11):1216.
[11]陈宏昌,魏文峰,霍金海,等.UPLC-Q-TOF-MS/MS分析刺五加叶的化学成分[J].中药材,2016,39(7):1536.
[12]迪丽拜尔·马木提.毛菊苣中秦皮甲素和秦皮乙素的药代动力学及生物转化研究[D].乌鲁木齐:新疆医科大学,2014.
[13] Ye M,Liu S H,Jiang Z L,et al. Liquid chromatography/mass spectrometry analysis of PHY906,a Chinese medicine formulation for cancertherapy[J]. Rapid Commun Mass Spectrom,2007,21(22):3593.
[14] Zhang L X,Zeng Y F,Yang X,et al. Chemical profiling of bioactive constituents in Sarcandra glabra and its preparations using ultra-high-pressure liquid chromatography coupled with LTQ Orbitrap mass spectrometry[J]. Rapid Commun Mass Spectrom,2011,25:2439.
[15]付珊.秦皮质量控制方法研究[D].石家庄:河北医科大学,2014.
[16]邓皓月,武媛媛,袁磊,等.基于UPLC-Q-TOF-MS分析不同产地抱茎苦荬菜化学成分的差异[J].中国实验方剂学杂志,2016,22(17):37.
[17] Shi Y,Wu C,Chen Y,et al. Comparative analysis of three Callicarpa herbs using high performance liquid chromatography with diode array detector and electrospray ionization-trap mass spectrometry method[J]. J Pharm Biomed Anal,2013,75(5):239.
[18]杨林军,谢彦云,李志锋,等.UPLC/Q-TOF-MS/MS分析中华常春藤中的化学成分[J].中草药,2016,47(4):566.
[19] Qi L W,Chen C Y,Li P,et al. Structureal characterization and identification of iridoid glycosides,sponins,phenolic acid and flavonoids in Flos Lonicerae Japonicea by a fast liquid chromatography method with diode-array detection and time-of-flight mass spectrometry[J]. Rapid Commun Mass Spectrom,2009,23(19):3227.
[20]何峰,王永林,郑林,等. UPLC-PDA-ESI-MS分析杜仲中化学成分[J].中国实验方剂学杂志,2014,20(3):59.
[21] Nishimura H,Sasaki H,Morota T,et al. Six glycosides from Rehmannia glutinosa var. purpurea[J]. Phytochemistry,1990,29(10):3303.
[22]杜憬生,吴立群,刘敬功,等.基于UPLC-Q-TOF-MS技术的菊花化学成分快速分析[J].中药材,2017,40(3):621.
[23]李泮霖,李楚源,刘孟华,等.基于UFLC-Triple-Q-TOF-MS/MS技术的金银花、山银花化学成分比较[J].中南药学,2016,14(4):363.
[24]李伟,冯育林,黎田儿,等. UPLC-Q-TOF/MS技术结合诊断离子方法快速分析连钱草中黄酮类化合物[J].质谱学报,2016,37(6):504.
[25]张靖,徐筱杰,徐文,等. HPLC-LTQ-Orbitrap-MSn快速鉴别党参药材中化学成分[J].中国实验方剂学杂志,2015,21(9):59.
[26] Fiorentino A,D'Abrosca B,Dellagreca M,et al. Chemical characterization of new oxylipins from Cestrum parqui, and their effects on seed germination and early seeding growth[J]. Chem Biodiv,2008,5(9):1780.
[27]李想.黄酮类化合物ESI-ITMSn质谱裂解规律的量子化学研究[D].佳木斯:佳木斯大学,2015.
[28]阿布拉江·克依木.黄酮苷类天然产物的质谱分析方法研究[D].北京:中国协和医科大学,2006.
[29]钟艳梅,冯毅凡,郭姣.基于UPLC-Q-TOF-MS技术的白术药材化学成分快速识别研究[J].质谱学报,2015,36(1):72.
[30]李蓉,秦坚民,余国莫.荨麻属药用植物研究概况及其开发价值[J].中国野生植物资源,2002,21(1):24.
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