基于LC-LTQ-Orbitrap/MS的复方木鸡颗粒成分分析
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摘要
目的:采用高效液相色谱-离子阱静电场轨道阱质谱(LC-LTQ-Orbitrap/MS)联用技术对满药复方木鸡颗粒甲醇提取物进行分析,确定其主要化学组分。方法:采用Waters Xselect T3 C_(18)色谱柱(250 mm×4.6 mm,5μm),以0.1%甲酸水(A)-乙腈(B)为流动相,梯度洗脱,流速0.5 mL·min~(-1),柱温30℃,进样量5μL;质谱采用电喷雾(ESI)离子源,在正负离子模式下采集数据,通过保留时间、精确相对分子质量和二级质谱碎片对检测到的化学成分进行鉴定,结合已有文献报道,确定化合物结构式。结果:鉴定或推断了复方木鸡颗粒甲醇提取物中36个主要化学成分,其中包括12个生物碱类化合物,13个黄酮类化合物,3个萘醌类化合物,6个酚酸类化合物和2个其他类化合物。结论:该方法快速、准确,为复方木鸡颗粒的化学成分鉴定提供一种新的策略,为复方木鸡颗粒药效物质基础和质量控制研究提供了实验基础。
Objective:To identify the major chemical constituents in compound Muji granules by liquid chromatography coupled with linear quadrupole ion trap-orbitrap mass spectrometry(LC-LTQ-Orbitrap/MS). Methods:The separation was performed on a Waters Xselect T3 C_(18) column(250 mm×4.6 mm,5 μm),and the mobile phase composed of 0.1% formic acid(A)and acetonitrile(B)with gradient elution at a flow rate of 0.5 mL·min~(-1). The column temperature was 30 ℃ and the injection volume was 5 μL. LTQ-Orbitrap/MS with electrospray ion(ESI)source was applied for the analysis under the positive and negative ion modes.The compounds were identified by retention time,accurate relative molecular mass and fragment ions in mass spectrometry combined with reference substances and references. Results:36 major chemical compounds including12 alkaloids,13 flavonoids,3 naphthalene quinones,6 phenolic acids and 2 other compounds were identified or inferred in compound Muji granules. Conclusion:The proposed method is fast and accurate,offering a new strategy for identification of the main chemical constituents in compound Muji granules,which can be used for quality evaluation and pharmacological research.
Objective:To identify the major chemical constituents in compound Muji granules by liquid chromatography coupled with linear quadrupole ion trap-orbitrap mass spectrometry(LC-LTQ-Orbitrap/MS). Methods:The separation was performed on a Waters Xselect T3 C_(18) column(250 mm×4.6 mm,5 μm),and the mobile phase composed of 0.1% formic acid(A)and acetonitrile(B)with gradient elution at a flow rate of 0.5 mL·min~(-1). The column temperature was 30 ℃ and the injection volume was 5 μL. LTQ-Orbitrap/MS with electrospray ion(ESI)source was applied for the analysis under the positive and negative ion modes.The compounds were identified by retention time,accurate relative molecular mass and fragment ions in mass spectrometry combined with reference substances and references. Results:36 major chemical compounds including12 alkaloids,13 flavonoids,3 naphthalene quinones,6 phenolic acids and 2 other compounds were identified or inferred in compound Muji granules. Conclusion:The proposed method is fast and accurate,offering a new strategy for identification of the main chemical constituents in compound Muji granules,which can be used for quality evaluation and pharmacological research.
引文
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[15]何常明.苦参和山豆根黄酮类成分及其生物活性的比较研究[D].上海:复旦大学,2010HE CM.Comparative Studies on Flavonoids and Their Bioactivities of Sophora flavescens and S.tokinensis[D].Shanghai:Fudan University,2010
[16]AHMADA A,TANDONA S,XUAN TD,et al.A review on phytoconstituents and biological activities of Cuscutaspecies[J].Biomed Pharmacother,2017,92:772
[17]李文兰,丁晶鑫,藏宝珊,等.基于HPLC-Q TOF-MS/MS技术分析纯化后菟丝子拟雌激素活性成分[J].中国药学杂志,2014,49(20):1791LI WL,DING JX,ZANG BS,et al.Analysis of purified active ingredients of estrogen mimic effect in Cuscuta chinensis by HPLC-QTOF-MS/MS[J].Chin Pharm J,2014,49(20):1791
[2]宋玉荣,姜春霞.满药复方木鸡颗粒研究新进展[J].中国医药导报,2013,10(1):4SONG YR,JIANG CX.New study progress of compound muji granule of Manchu medicine[J].China Med Her,2013,10(1):4
[3]时晓亚,李志浩,李鹏.HPLC法测定复方木鸡颗粒中金丝桃苷的含量[J].中国医药导报,2015,21(20):28SHI XY,LI ZH,LI P.Determination of hyperoside in Fufang Muji granules by HPLC[J].China Med Her,2015,21(20):28
[4]郑阳阳,王帅,孟宪生,等.复方木鸡颗粒对小鼠慢性肝损伤的影响[J].亚太传统医药,2016,12(5):13ZHENG YY,WANG S,MENG XS,et al.The influence of compound muji granules on chronic liver in injury in mice[J].Asia-Pacific Tradit Med,2016,12(5):13
[5]黄海燕,鲁静.复方木鸡颗粒检测方法研究[J].药物分析杂志,2007,27(8):1190HUANG HY,LU J.Study on the quality control method of compound Muji granules[J].Chin J Pharm Anal,2007,27(8):1190
[6]王颖,王帅,包永睿,等.复方木鸡颗粒对CCl4诱导大鼠肝硬化的防治作用[J].中成药,2017,39(3):597WANG Y,WANG S,BAO YR,et al.The compound Muji granules on prevention and treatment of liver cirrhosis in rats induced by CCl4[J].Chin Tradit Pat Med,2017,39(3):597
[7]孙俊艳.雄激素受体信号通路对神经母细胞瘤的调节作用及复方木鸡颗粒对神经母细胞瘤治疗作用的初步观察[D].大连:大连医科大学,2017SUN JY.Androgen Receptor Signaling Regulates the Growth of Neuroblastoma Cells In Vitro and In Vivo and Initial Observation of the Effect of Fufangmujikeli on Neuroblastoma[D].Dalian:Dalian Medical University,2017
[8]程钱.山豆根指纹图谱、含量测定及成分分析研究[D].北京:北京中医药大学,2017CHENG Q.Studies on Fingerprint,Determination of the Content and Composition Analysis of Sophorae tonkinensis[D].Beijing:Beijing university of Chinese Medicine,2017
[9]郭智.基于UPLC-ESI-QTof的山豆根、苦参化学成分比较及柴胡不同提取方法柴胡皂苷a、d变化的研究[D].北京:北京协和医学院,2013GUO Z.Studies on Chemical Constituents of S.tonkinensis and S.flavescens and Transformation of Saikosaponin a,d in Different Extracting Method by Using UPLC-ESI-Qtof[D].Beijing:Peking Union Medical College,2013
[10]曾祖平,郭智,彭冰,等.山豆根和苦参碱类成分UPLC/Q-TOFMSE比较研究[J].天然产物研究与开发,2015,27(5):804ZENG ZP,GUO Z,PENG B,et al.Comparative analysis of alkaloids in Radix Sophorae Tonkinensis and Radix Sophorae Flavescentis using UPLC/Q-TOF MSE[J].Nat Prod Res Dev,2015,27(5):804
[11]王增绘,王冬梅,刘艾琳,等.山豆根抑制丁酰胆碱酯酶活性及活性部位UPLC-Q-TOF-MS分析[J].中国现代中药,2015,17(9):912WANG ZH,WANG DM,LIU AL,et al.Analysis of chemical constituents of effective part of anti-butyrylcholinesterase of Sophora tonkinensis by UPLC-Q-TOF-MS[J].Mod Chin Med,2015,17(9):912
[12]陈怀侠,韩凤梅,杜鹏,等.苦参生物碱电喷雾质谱分析[J].分析化学,2006,34(2):205CHEN HX,HAN FM,DU P,et al.Electrospray ionization mass spectrometry for analyses of alkaloids in Sophora flavescens Ait[J].Chin J Anal Chem,2006,34(2):205
[13]ZOU DX,WANG JF,ZHANG B,et al.Analysis of chemical constituents in Wuzi-Yanzong-Wan by UPLC-ESI-LTQ-OrbitrapMS[J].Molecules,2015,20(12):21373
[14]孙国东,霍金海,王改丽,等.基于UPLC-Q-TOF/MS技术的核桃楸皮成分分析[J].中草药,2017,48(4):657SUN GD,HUO JH,WANG GL,et al.Identification and characterization of chemical constituents in Cortex Juglandis Mandshuricae based on UPLC-Q-TOF/MS[J].Chin Tradit Herb Drugs,2017,48(4):657
[15]何常明.苦参和山豆根黄酮类成分及其生物活性的比较研究[D].上海:复旦大学,2010HE CM.Comparative Studies on Flavonoids and Their Bioactivities of Sophora flavescens and S.tokinensis[D].Shanghai:Fudan University,2010
[16]AHMADA A,TANDONA S,XUAN TD,et al.A review on phytoconstituents and biological activities of Cuscutaspecies[J].Biomed Pharmacother,2017,92:772
[17]李文兰,丁晶鑫,藏宝珊,等.基于HPLC-Q TOF-MS/MS技术分析纯化后菟丝子拟雌激素活性成分[J].中国药学杂志,2014,49(20):1791LI WL,DING JX,ZANG BS,et al.Analysis of purified active ingredients of estrogen mimic effect in Cuscuta chinensis by HPLC-QTOF-MS/MS[J].Chin Pharm J,2014,49(20):1791