β_1-AR/CMC-UPLC/MS联用的分析方法筛选辛夷中的活性成分
|
摘要
目的建立高表达的β_1肾上腺素受体细胞膜色谱法(β_1-AR/CMC)与超高效液相-质谱(UPLC/MS)离线联用(β_1-AR/CMC-Offline-UPLC/MS)的分析方法,并从辛夷中筛选对β_1-AR具有抑制作用的活性成分。方法建立β_1-AR/CMC-Offline-UPLC/MS分析方法;采用阳性对照药物美托洛尔(Meto)和阴性对照药物地西泮(DZ)考察分析方法的可行性;用所建立的分析方法分离、筛选、鉴定辛夷提取物中的活性成分。结果建立并验证了β_1-AR/CMC-Offline-UPLC/MS分析方法,并以β_1-AR/CMC系统对辛夷95%乙醇提取物中的活性成分进行筛选,经UPLC/MS鉴定保留组分为辛夷脂素。结论本课题所建立的β_1-AR/CMC-Offline-UPLC/MS分析方法具有灵敏度高、特异性强、稳定性高、分析速度快的特点,能够用于从辛夷中筛选对β1-AR具有抑制作用的活性成分。
Objective To develop a highly expressed β_1-AR cell membrane chromatography(β_1-AR/CMC) offline coupling ultra-performance liquid chromatography and mass spectrometry(UPLC/MS) method for screening active ingredients from Magnoliae Flos.Methods β_1-AR/CMC-Offline-UPLC/MS analytical method was established. A negative control drug diazepam(DZ) and a positive control drug metoprolol were injected into the β_1-AR/CMC column to determine the validation. The active ingredients in extracts of Magnoliae Flos were separated and screened using the β_1-AR/CMC system,and then the obtained fractions were identified by UPLC/MS system. Results The β_1 AR/CMC-Offline-UPLC/MS method was verified. The active ingredients were screened from 95% ethanol extract of Magnoliae Flos on the β_1-AR/CMC,and the obtained fraction was identified as fargesin by UPLC/MS. Conclusion The established β_1-AR/CMC-offline-UPLC/MS method could be used as a sensitive,specific,stable,and quick alternative method for screening new type of β_1-AR antagonists from Magnoliae Flos.
Objective To develop a highly expressed β_1-AR cell membrane chromatography(β_1-AR/CMC) offline coupling ultra-performance liquid chromatography and mass spectrometry(UPLC/MS) method for screening active ingredients from Magnoliae Flos.Methods β_1-AR/CMC-Offline-UPLC/MS analytical method was established. A negative control drug diazepam(DZ) and a positive control drug metoprolol were injected into the β_1-AR/CMC column to determine the validation. The active ingredients in extracts of Magnoliae Flos were separated and screened using the β_1-AR/CMC system,and then the obtained fractions were identified by UPLC/MS system. Results The β_1 AR/CMC-Offline-UPLC/MS method was verified. The active ingredients were screened from 95% ethanol extract of Magnoliae Flos on the β_1-AR/CMC,and the obtained fraction was identified as fargesin by UPLC/MS. Conclusion The established β_1-AR/CMC-offline-UPLC/MS method could be used as a sensitive,specific,stable,and quick alternative method for screening new type of β_1-AR antagonists from Magnoliae Flos.
引文
[1]陈蓁,王锐平,贺浪冲,等.细胞膜色谱法筛选红花中的有效成分[J].陕西中医,2004,25(7):643-644.
[2]管政,马小卓,吕圭源,等.辛夷挥发油对变应性鼻炎大鼠IL-12\IFN-γ及组胺的影响[J].中药药理与临床,2011,27(2):70-72.
[3]刘小丽,邢晓为,黄利华,等.β1肾上腺素受体与心衰、高血压和心肌纤维化之间的关系研究进展[J].中国药理学通报,2013,45(12):1640-1644.
[4]杨兴鑫,张艳利,李晓妮.细胞膜色谱法及其在药物研究中的应用进展[J].中国药学杂志,2010,45(8):361-1364.
[5]王新,薛晖,岳媛,等.高表达β1肾上腺素受体细胞膜色谱法靶向筛选夏天无中活性成分[J].中国实验方剂学杂志,2015,18(21):65-68.
[6] Yue Y,Xue H,Wang X,et al. High-expressionβ1adrenergic receptor/cell membrane chromatography method based on a target receptor to screen active ingredients from traditional Chinese medicines[J]. J Sep Sci,2014,37(3). 244-249.
[7] Zhang Y,Cao YJ,Zhan YZ,et al. Furanocoumarins-imperatorin inhibits myocardial hypertrophy both in vitro and in vivo[J]. Fitoterapia,2010,81:1188-1195.
[8]冯卫生,王建超,何玉环,等.辛夷化学成分的研究[J].中国药学杂志,2015,50(24):2103-2106.
[9]王永慧,叶方,张秀华.辛夷药理作用和临床应用研究进展[J].中国医药导报,2012,9(16):12-14.
[10]王甜甜曹赟蒋运斌,等.中药辛夷研究进展[J].亚太传统医药,2017,13(18):74-78.
[11] Hou XF,Zhou MZ,Jiang Q,et al. A vascular smooth muscle/cell membrane chromatography-offline-gas chromatography/mass spectrometry method for recognition,separation and identification of active components from traditional Chinese medicines[J]. J Chromatogr A,2009,1216:7081-7087.
[12] Chang RM,Li Y,Yang XX,et al. Protective role of Deoxyschizandrin and Schisantherin A against myocardial ischemia-reperfusion injury in rats[J]. PLo S One,2013,8:e61590.
[2]管政,马小卓,吕圭源,等.辛夷挥发油对变应性鼻炎大鼠IL-12\IFN-γ及组胺的影响[J].中药药理与临床,2011,27(2):70-72.
[3]刘小丽,邢晓为,黄利华,等.β1肾上腺素受体与心衰、高血压和心肌纤维化之间的关系研究进展[J].中国药理学通报,2013,45(12):1640-1644.
[4]杨兴鑫,张艳利,李晓妮.细胞膜色谱法及其在药物研究中的应用进展[J].中国药学杂志,2010,45(8):361-1364.
[5]王新,薛晖,岳媛,等.高表达β1肾上腺素受体细胞膜色谱法靶向筛选夏天无中活性成分[J].中国实验方剂学杂志,2015,18(21):65-68.
[6] Yue Y,Xue H,Wang X,et al. High-expressionβ1adrenergic receptor/cell membrane chromatography method based on a target receptor to screen active ingredients from traditional Chinese medicines[J]. J Sep Sci,2014,37(3). 244-249.
[7] Zhang Y,Cao YJ,Zhan YZ,et al. Furanocoumarins-imperatorin inhibits myocardial hypertrophy both in vitro and in vivo[J]. Fitoterapia,2010,81:1188-1195.
[8]冯卫生,王建超,何玉环,等.辛夷化学成分的研究[J].中国药学杂志,2015,50(24):2103-2106.
[9]王永慧,叶方,张秀华.辛夷药理作用和临床应用研究进展[J].中国医药导报,2012,9(16):12-14.
[10]王甜甜曹赟蒋运斌,等.中药辛夷研究进展[J].亚太传统医药,2017,13(18):74-78.
[11] Hou XF,Zhou MZ,Jiang Q,et al. A vascular smooth muscle/cell membrane chromatography-offline-gas chromatography/mass spectrometry method for recognition,separation and identification of active components from traditional Chinese medicines[J]. J Chromatogr A,2009,1216:7081-7087.
[12] Chang RM,Li Y,Yang XX,et al. Protective role of Deoxyschizandrin and Schisantherin A against myocardial ischemia-reperfusion injury in rats[J]. PLo S One,2013,8:e61590.