基于UPLC-Q-Exactive四极杆-静电场轨道阱高分辨质谱联用技术快速识别补肾活血方的化学成分
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摘要
目的:利用UPLC-Q-Exactive四极杆-静电场轨道阱高分辨质谱仪联用技术,快速识别与鉴定补肾活血方中的化学成分。方法:采用ACQUITY UPLC BEH C_(18)色谱柱(100 mm×2.1 mm,1.7μm),以乙腈-0.1%甲酸水溶液为流动相进行梯度洗脱,质谱采用负离子监测模式、全扫描及自动触发二级质谱扫描功能,对补肾活血方化学成分进行快速识别及鉴定。结果:通过质谱数据、对照品比对共鉴定出58个化合物;结果显示,补肾活血方中的主要化学成分包括异黄酮类、酚酸类、人参皂苷类及少量的环烯醚萜类化合物。结论:该研究能够快速分析和确定补肾活血方的化学成分,为确定补肾活血方防治糖尿病性视网膜病变药效物质提供了依据。
Objective:To develop an ultra-high performance liquid chromatography couple with Q-Exactive quadrupole-Orbitrap mass spectrometry method for rapid recognition and identification of the chemical constituents from Bushen Huoxue prescription.Methods:The Bushen Huoxue prescription extract was separated on an ACQUITY UPLC BEH C_(18) column(100 mm×2.1 mm,1.7 μm),and eluted with a gradient of methanol-water containing 0.1% formic acid.The constituents from Bushen Huoxue prescription were rapidly recognized and identified by HRMS in the negative ion mode using both full scan and automatic triggering of the function of the two stage mass spectrometry scanning.Results:A total of 58 compounds were identified by referring to mass spectrometric data and comparing with reference substances.Bushen Huoxue prescription mainly contains isoflavones,phenolic acids,ginsenosides and a small quantity of iridoids.Conclusion:The research can be used to quickly analyze and determine chemical constituents from Bushen Huoxue prescription,which also provides an experimental basis for further study of effective components in the prevention and treatment of diabetic retinopathy.
Objective:To develop an ultra-high performance liquid chromatography couple with Q-Exactive quadrupole-Orbitrap mass spectrometry method for rapid recognition and identification of the chemical constituents from Bushen Huoxue prescription.Methods:The Bushen Huoxue prescription extract was separated on an ACQUITY UPLC BEH C_(18) column(100 mm×2.1 mm,1.7 μm),and eluted with a gradient of methanol-water containing 0.1% formic acid.The constituents from Bushen Huoxue prescription were rapidly recognized and identified by HRMS in the negative ion mode using both full scan and automatic triggering of the function of the two stage mass spectrometry scanning.Results:A total of 58 compounds were identified by referring to mass spectrometric data and comparing with reference substances.Bushen Huoxue prescription mainly contains isoflavones,phenolic acids,ginsenosides and a small quantity of iridoids.Conclusion:The research can be used to quickly analyze and determine chemical constituents from Bushen Huoxue prescription,which also provides an experimental basis for further study of effective components in the prevention and treatment of diabetic retinopathy.
引文
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[3]谢学军,李翔,肖丹,等.补肾活血中药对糖尿病大鼠视网膜神经节细胞的影响[J].中国中医眼科杂志,2000,10(4),195XIE XJ,LI X,XIAO D,et al.An observation of influence of tradition Chinese medicine for Bushenhuoxue therapy on retinal ganglion cells in experimental diabetic rats[J].J Tradit Chin Ophthalmol,2000,10(4):195
[4]谢学军,宋明霞,张梅,等.补肾活血中药复方对高糖及糖基化终末产物条件下视网膜Müller细胞的影响[J].中国中西医结合杂志,2015,35(6):735XIE XJ,SONG MX,ZHANG M,et al.Effect of Bushenhuoxue compound on retinal MüllercellsinhighglucoseorAGEs conditions[J].Chin J Integr Tradit West Med,2015,35(6):735
[5]秦伟,谢学军,何宇,等.补肾活血中药对糖尿病大鼠视路Nissl小体和Bcl-2的影响[J].江西中医学院学报,2011,23(6):65QIN W,XIE XJ,HE Y,et al.The effect of Chinese medicine of Bushenhuoxue therapy on Nissl bodies and B cell lymphoma/leukemia-2 in the visual pathway of experimental diabetic rats[J].J Jiangxi Univ TCM,2011,23(6):65
[6]马荣,谢学军,万李,等.补肾活血中药血清对高糖条件下纯化培养的视网膜神经节细胞Glu释放量的影响[J].中国中西医结合杂志,2010,30(8):875MA R,XIE XJ,WAN L,et al.Effect of drug-serum with Chinese drugs for nourishing Shen and activating blood on glutamate release in purified retinal ganglion cells cultured in high-glucose conditions[J].Chin J Integr Tradit West Med,2010,30(8):875
[7]谢学军,张梅,王明芳.一种中药组合物及其制备方法:中国,CN200910308470.3[P].2010-03-03XIE XJ,ZHANG M,WANG MF.A Kind of Traditional Chinese Medicine Combination and Its Preparation Method:China,200910308470.3[P].2010-03-03
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[11]赵新峰,李平,孙毓庆.熟地黄的高效液相色谱/电喷雾电离-质谱分析[J].药物分析杂志,2007,27(6):874ZHAO XF,LI P,SUN YQ.HPLC/ESI-MS analysis of the Radix Rehmanniae Praeprata[J].Chin J Pharm Anal,2007,27(6):874
[12]陈慧贞,李云飞,钟敬华,等.脑得生片物质基础的HPLC-DADMSn分析[J].浙江大学学报(医学版),2012,41(1):32CHEN HZ,LI YF,ZHONG JH,et al.Identification of major component of traditional Chinese medicine Naodesheng tablets by HPLC-DADMSn[J].J Zhejiang Univ(Med Sci),2012,41(1):32
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[17]TAWAB MA,BAHR U,KARAS M,et al.Degradation of ginsenosides in humans after oral administration[J].Drug Metab Dispos,2003,31(8):1065
[18]TANSAKUL P,SHIBUYA M,KUSHIRO T,et al.Dammarenediol-Ⅱsynthase,the first dedicated enzyme for ginsenoside biosynthesis,in Panax ginseng[J].FEBS Lett,2006,580(22):5143
[19]SUN BS,XU MY,LI Z,et al.UPLC-Q-TOF-MS/MS analysis for steaming times-dependent profiling of steamed Panax quinquefolius and its ginsenosides transformations induced by repetitious steaming[J].J Ginseng Res,2012,36(3):277
[20]周思思,马增春,梁乾德,等.基于UPLC-TOF-MS分析人参麦冬配伍后人参皂苷类成分的变化[J].质谱学报,2013,34(2):88ZHOU SS,MA ZC,LIANG QD,et al.UPLC-TOF-MS based profilingapproachtoevaluateginsenosidecompositionin combination of Ginseng and Radix Ophiopogonis[J].J Chin Mass Spectrom Soc,2013,34(2):88
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