UPLC-Q-TOF-MS分析补肾强身片中化学成分
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摘要
目的:采用超高效液相色谱-四级杆-飞行时间质谱(UPLC-Q-TOF-MS)对补肾强身片中的化学成分进行研究。方法:以Agilent SB-C18色谱柱(4.6 mm×100 mm,2.7μm),流动相乙腈-0.1%甲酸水溶液,梯度洗脱,流速0.5 m L·min~(-1),柱温40℃,电喷雾离子源,正、负离子模式下采集数据。首先对有对照品的成分进行定性分析,对于多数没有对照品的成分,主要根据UPLC-Q-TOF-MS提供的准确相对分子质量和离子碎片信息并结合文献数据比对进行化学成分结构推测鉴定。结果:共鉴定了补肾强身片中50个化学成分,包括酚酸类19个、黄酮类25个、裂环环烯醚萜类4个、苯乙醇苷类1个、生物碱类1个,其中12个成分(原儿茶酸,红景天苷,原儿茶醛,绿原酸,金丝桃苷,异槲皮苷,特女贞苷,朝藿定C,淫羊藿苷,山柰酚,宝霍苷I,紫云英苷)通过与对照品比对进行定性分析。结论:UPLC-Q-TOF-MS可以简便、快速地对补肾强身片中化学成分定性分析,该研究丰富了补肾强身片中化学成分的信息,为补肾强身片药效物质基础研究、质量控制及其临床用药提供了参考。
Objective: To investigate the chemical constituents in Bushen Qiangshen pills( BQP) by ultra-performance liquid chromatography-quadrupole-time of flight mass spectrometry( UPLC-Q-TOF-MS).Method: The UPLC separation was performed on Agilent SB-C18 reversed-phase column( 4. 6 mm × 100 mm,2. 7 μm) with acetonitrile-0. 1% formic acid solution as mobile phase for gradient elution. The flow rate was0. 5 m L·min~(-1),and the column temperature was 40 ℃. Data were collected via positive and negative ion mode using ESI ion source. First of all,the qualitative analysis was conducted for the components with reference substances,and the major components without reference substances were identified according to the relative molecule weight and fragment information by UPLC-Q-TOF-MS as well as comparison with literature data. Result:Fifty compounds from BQP were identified, including 19 phenolic acids, 25 flavonoids, 4 olefins, 1 phenylethanolate and 1 alkaloid. 12 components( protocatechuic acid, salidroside, protocatechuic aldehyde,chlorogenic acid, hyperin, isoquercitrin, specnuezhenide, epimedin C, icariin, kaempferide,baohuoside Ⅰand astragalin) among them were qualitatively analyzed by comparing with reference substances. Conclusion:UPLC-Q-TOF-MS is convenient and rapid to analyze the chemical constituents in BQP,providing a reference for the study on material foundation of efficacy,quality control and clinical application of BQP.
Objective: To investigate the chemical constituents in Bushen Qiangshen pills( BQP) by ultra-performance liquid chromatography-quadrupole-time of flight mass spectrometry( UPLC-Q-TOF-MS).Method: The UPLC separation was performed on Agilent SB-C18 reversed-phase column( 4. 6 mm × 100 mm,2. 7 μm) with acetonitrile-0. 1% formic acid solution as mobile phase for gradient elution. The flow rate was0. 5 m L·min~(-1),and the column temperature was 40 ℃. Data were collected via positive and negative ion mode using ESI ion source. First of all,the qualitative analysis was conducted for the components with reference substances,and the major components without reference substances were identified according to the relative molecule weight and fragment information by UPLC-Q-TOF-MS as well as comparison with literature data. Result:Fifty compounds from BQP were identified, including 19 phenolic acids, 25 flavonoids, 4 olefins, 1 phenylethanolate and 1 alkaloid. 12 components( protocatechuic acid, salidroside, protocatechuic aldehyde,chlorogenic acid, hyperin, isoquercitrin, specnuezhenide, epimedin C, icariin, kaempferide,baohuoside Ⅰand astragalin) among them were qualitatively analyzed by comparing with reference substances. Conclusion:UPLC-Q-TOF-MS is convenient and rapid to analyze the chemical constituents in BQP,providing a reference for the study on material foundation of efficacy,quality control and clinical application of BQP.
引文
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[2]贾福华.补肾强身片的临床治验[J].中成药研究,1982(8):28.
[3]王继玉,卑国斌.改进补肾强身片淫羊藿苷薄层色谱鉴别方法的建议[J].辽宁中医杂志,2010,37(S1):207-208.
[4]孙明,郑爱华.HPLC测定补肾强身片中淫羊藿苷的含量[J].中成药,2006,28(8):1231-1232.
[5]方慧祥,杨坤芬,冯玉茹.RP-HPLC法测定补肾强身片中金丝桃苷的含量[J].中国药房,2015,26(3):413-415.
[6]穆天慧,方慧祥,黎海存.HPLC法测定补肾强身片中原儿茶酸的含量[J].中国民族民间医药,2014,23(9):13-14.
[7]雷勇胜,宋丽明,蒋庆峰.液质联用技术在药物的有关物质分析中的应用[J].现代仪器,2011,17(4):9-13,17.
[8]Clifford M N,Johnston K L,Knight S,et al.Hierarchical scheme for LC-MSnidentification of chlorogenic acids[J].J Agr Food Chem,2003,51(10):2900-2911.
[9]李辰辰,陆小腾驾,童华荣.HPLC-Q-TOF-MS-MS测定桑椹中多酚类物质[J].食品科学,2015,36(2):101-104.
[10]XU J S,YANG L,ZENG X,et al.Characterization of compounds in the Chinese herbal drug Mu-Dan-Pi by liquid chromatography coupled to electrospray ionization mass spectrometry[J].Rapid Commun Mass Spectrom,2006,20(22):3275-3288.
[11]ZHANG J Y,ZHANG Q,LI N,et al.Diagnostic fragment-ion-based and extension strategy coupled to DFIs intensity analysis for identification of chlorogenic acids isomers in Flos Lonicerae Japonicae by HPLC-ESIMSn[J].Talanta,2013,104:1-9.
[12]刘颖,卢建秋,张加余,等.苦碟子药材及其注射液中化学成分的HPLC-ESI-MSn分析[J].中国中药杂志,2013,38(16):2675-2681.
[13]谭乐俊,王萌,任晓亮,等.基于UPLC-Q/TOF-MS的中药水蜜丸制剂二至丸多成分稳定性研究[J].中草药,2016,47(13):2260-2268.
[14]Parveen I,Threadgill M D,Hauck B,et al.Isolation,identification and quantitation of hydroxycinnamic acid conjugates,potential platform chemicals,in the leaves and stems of Miscanthus×giganteus using LC-ESI-MSn[J].Phytochemistry,2011,72(18):2376-2384.
[15]王放,张加余,王青,等.HPLC-HR-MSn法鉴定苦碟子注射液中酚酸类成分[J].中南药学,2013,11(8):561-565.
[16]叶敏,阎玉凝,乔梁,等.中药菟丝子化学成分研究[J].中国中药杂志,2002,27(2):115-117.
[17]Jaiswal R,Kiprotich J,Kuhnert N.Determination of the hydroxycinnamate profile of 12 members of the Asteraceae family[J].Phytochemistry,2011,72(8):781-790.
[18]ZHOU J,SUN J B,ZHENG P,et al.Orthogonal array design for optimization of hollow-fiber-based liquidphase microextraction combined with high-performance liquid chromatography for study of the pharmacokinetics of magnoflorine in rat plasma[J].Anal Bioanal Chem,2012,403(7):1951-1960.
[19]ZHAO H Y,SUN J H,FAN M X,et al.Analysis of phenolic compounds in Epimedium plants using liquid chromatography coupled with electrospray ionization mass spectrometry[J].J Chromatogr A,2008,1190(1/2):157-181.
[20]宋梦晗,张学兰,李慧芬,等.LC-TOF/MS快速鉴定女贞子4种环烯醚萜苷类化合物模拟清蒸品的化学成分[J].中国实验方剂学杂志,2016,22(16):14-17.
[21]LI H F,GUAN X Y,YE M,et al.Qualitative and quantitative analyses of Epimedium wushanense by highperformance liquid chromatography coupled with diode array detection and electrospray ionization tandem mass spectrometry[J].J Separation Sci,2015,34(12):1437-1446.
[22]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):37-42.
[23]袁航.黔淫羊藿的鉴定及质量评价研究[D].北京:北京中医药大学,2014.
[24]舒俊生,黄桂东,毛健.滁菊95%乙醇提取液中黄酮类和酚酸类物质的鉴定及分离[J].中国食品学报,2013,13(4):207-213.
[25]袁杰,龚又明,鞠鹏,等.HPLC-MS2法分析朝鲜淫羊藿中的化学成分[J].中草药,2004,35(4):371-374.
[26]LI H F,GUAN X Y,YE M,et al.Qualitative and quantitative analyses of Epimedium wushanense by highperformance liquid chromatography coupled with diode array detection and electrospray ionization tandem mass spectrometry[J].J Sep Sci,2011,34(12):1437-1446.