UPLC-DAD-MS定性和定量分析蕲艾中的酚酸和黄酮类成分
|
摘要
采用UPLC-LTQ-Orbitrap-MS鉴定蕲艾中的酚酸和黄酮类成分,UPLC-DAD测定药材中主要成分的含量。采用Xtimate UHPLC C18色谱柱(2. 1 mm×100 mm,1. 8μm),以乙腈-0. 1%甲酸水溶液梯度洗脱,流速0. 4 m L·min-1,检测波长330nm,质谱扫描为正离子模式。通过质谱数据解析、对照品和文献对照,共鉴定、推导出蕲艾中6个酚酸和10个黄酮类成分,并建立了新绿原酸、绿原酸、隐绿原酸、异绿原酸A、异绿原酸B、异绿原酸C、茵陈色原酮、高车前素、棕矢车菊素、5,7,3'-三羟基-6,4',5'-三甲氧基黄酮、异泽兰黄素和蔓荆子黄素12个成分的含量测定方法,各成分质量浓度与峰面积具有较好的线性关系;精密度、重复性和稳定性良好(RSD<3. 0%);加样回收率为103. 3～109. 2%。该方法新颖、快速、准确,可作为蕲艾全面可靠的质量控制方法,应用于不同产地艾叶中相关成分的测定,发现蕲艾中的成分含量具有独特性。
The study aims to qualitatively and quantitatively analyze phenolic acids and flavonoids in Artemisiae Argyi Folium cultivated in Qichun(Qiai) for the quality control of this genuine regional herbs. UPLC-LTQ-Orbitrap-MS was used for rapid separation and structural identification of the constituents. Samples were separated on an UPLC column(2. 1 mm×100 mm,1. 8 μm) by gradient elution using 0. 1% formic acid and acetonitrile as mobile phases at a flow rate of 0. 4 m L·min-1. By UPLC-LTQ-Orbitrap-MS,16 compounds including phenolic acids and flavonoids were identified by comparison with reference standards or literature data. For quantitative analysis,12 identified compounds were simultaneously determined by UPLC-DAD at wavelengths of 330 nm. The method was validated with respect to linearity,precision,repeatability,stability and recovery. The contents of these compounds were found to differ significantly between the samples from Qichun and other areas. This strategy was novel,effective and straightforward,which provided a potential approach for holistic quality control of Qiai.
The study aims to qualitatively and quantitatively analyze phenolic acids and flavonoids in Artemisiae Argyi Folium cultivated in Qichun(Qiai) for the quality control of this genuine regional herbs. UPLC-LTQ-Orbitrap-MS was used for rapid separation and structural identification of the constituents. Samples were separated on an UPLC column(2. 1 mm×100 mm,1. 8 μm) by gradient elution using 0. 1% formic acid and acetonitrile as mobile phases at a flow rate of 0. 4 m L·min-1. By UPLC-LTQ-Orbitrap-MS,16 compounds including phenolic acids and flavonoids were identified by comparison with reference standards or literature data. For quantitative analysis,12 identified compounds were simultaneously determined by UPLC-DAD at wavelengths of 330 nm. The method was validated with respect to linearity,precision,repeatability,stability and recovery. The contents of these compounds were found to differ significantly between the samples from Qichun and other areas. This strategy was novel,effective and straightforward,which provided a potential approach for holistic quality control of Qiai.
引文
[1]李真真,吕洁丽,张来宾,等.艾叶的化学成分及药理作用研究进展[J].国际药学研究杂志,2016,43(6):1059.
[2]洪宗国.蕲艾的道地性研究[J].中南民族大学学报:自然科学版,2015,34(2):33.
[3]曹玲,于丹,崔磊,等.艾叶的化学成分、药理作用及产品开发研究进展[J].药物评价研究,2018,41(5):918.
[4] Han B,Xin Z,Ma S,et al. Comprehensive characterization and identification of antioxidants in Folium Artemisiae Argyi using high-resolution tandem mass spectrometry[J]. J Chromatogr B Analyt Technol Biomed Life Sci,2017,1063:84.
[5] Justesen U. Collision-induced fragmentation of deprotonated methoxylated flavonoids,obtained by electrospray ionization mass spectrometry[J]. J Mass Spectrom,2001,36(2):169.
[6]王燕,李晓波,李俊杰,等. 5,6,7,4'-四羟基黄酮和5,6,7,4'-四甲氧基黄酮高分辨电喷雾串联质谱裂解规律对比研究[J].质谱学报,2016,37(5):385.
[7]中国药典.一部[S]. 2015:83.
[8]刘红杰,白杨,洪燕龙,等.不同提取方法制备的艾叶挥发油化学成分分析与急性肝毒性比较[J].中国中药杂志,2010,35(11):1439.
[9]许俊洁,卢金清,郭胜男,等.不同部位与不同采收期蕲艾精油化学成分的GC-MS分析[J].中国实验方剂学杂志,2015,21(21):51.
[10]吴娜,孙智达.艾蒿黄酮体外抗氧化活性及对DNA氧化损伤的保护研究[J].食品科学,2008,29(10):47.
[11] Jeong D,Yi Y S,Sung G H,et al. Anti-inflammatory activities and mechanisms of Artemisia asiatica ethanol extract[J]. J Ethnopharmacol,2014,152(3):487.
[12]郭龙,焦倩,张丹,等.基于指纹图谱和多组分含量测定的艾叶药材质量控制研究[J].中国中药杂志,2018,43(5):977.
[13]王宇卿,庄果,张须学,等.多元统计分析比较宛艾及31个产地艾叶HPLC指纹图谱[J].中成药,2018,40(3):653.
[14]陈小露,梅全喜.艾叶化学成分研究进展[J].今日药学,2013,23(12):848.
[15]刘梦菲,江汉美,肖宇硕,等.HS-SPME-GC-MS联用技术分析不同产地艾叶挥发性成分[J].中国实验方剂学杂志,2018,24(10):79.
[16]王远敏,吴红梅,杨烨,等.艾叶UPLC指纹图谱[J].中国实验方剂学杂志,2015,21(13):50.
[17]徐文,丘小惠,张靖,等.超高压液相/电喷雾-LTQ-Orbitrap质谱联用技术分析三七根中皂苷类成分[J].药学学报,2012,47(6):773.
[18] Mei Q,Chen X,Xiang L,et al. DNA Barcode for identifying Folium Artemisiae Argyi from counterfeits[J]. Biol Pharm Bull,2016,39(9):1531.
[2]洪宗国.蕲艾的道地性研究[J].中南民族大学学报:自然科学版,2015,34(2):33.
[3]曹玲,于丹,崔磊,等.艾叶的化学成分、药理作用及产品开发研究进展[J].药物评价研究,2018,41(5):918.
[4] Han B,Xin Z,Ma S,et al. Comprehensive characterization and identification of antioxidants in Folium Artemisiae Argyi using high-resolution tandem mass spectrometry[J]. J Chromatogr B Analyt Technol Biomed Life Sci,2017,1063:84.
[5] Justesen U. Collision-induced fragmentation of deprotonated methoxylated flavonoids,obtained by electrospray ionization mass spectrometry[J]. J Mass Spectrom,2001,36(2):169.
[6]王燕,李晓波,李俊杰,等. 5,6,7,4'-四羟基黄酮和5,6,7,4'-四甲氧基黄酮高分辨电喷雾串联质谱裂解规律对比研究[J].质谱学报,2016,37(5):385.
[7]中国药典.一部[S]. 2015:83.
[8]刘红杰,白杨,洪燕龙,等.不同提取方法制备的艾叶挥发油化学成分分析与急性肝毒性比较[J].中国中药杂志,2010,35(11):1439.
[9]许俊洁,卢金清,郭胜男,等.不同部位与不同采收期蕲艾精油化学成分的GC-MS分析[J].中国实验方剂学杂志,2015,21(21):51.
[10]吴娜,孙智达.艾蒿黄酮体外抗氧化活性及对DNA氧化损伤的保护研究[J].食品科学,2008,29(10):47.
[11] Jeong D,Yi Y S,Sung G H,et al. Anti-inflammatory activities and mechanisms of Artemisia asiatica ethanol extract[J]. J Ethnopharmacol,2014,152(3):487.
[12]郭龙,焦倩,张丹,等.基于指纹图谱和多组分含量测定的艾叶药材质量控制研究[J].中国中药杂志,2018,43(5):977.
[13]王宇卿,庄果,张须学,等.多元统计分析比较宛艾及31个产地艾叶HPLC指纹图谱[J].中成药,2018,40(3):653.
[14]陈小露,梅全喜.艾叶化学成分研究进展[J].今日药学,2013,23(12):848.
[15]刘梦菲,江汉美,肖宇硕,等.HS-SPME-GC-MS联用技术分析不同产地艾叶挥发性成分[J].中国实验方剂学杂志,2018,24(10):79.
[16]王远敏,吴红梅,杨烨,等.艾叶UPLC指纹图谱[J].中国实验方剂学杂志,2015,21(13):50.
[17]徐文,丘小惠,张靖,等.超高压液相/电喷雾-LTQ-Orbitrap质谱联用技术分析三七根中皂苷类成分[J].药学学报,2012,47(6):773.
[18] Mei Q,Chen X,Xiang L,et al. DNA Barcode for identifying Folium Artemisiae Argyi from counterfeits[J]. Biol Pharm Bull,2016,39(9):1531.