葛根的指纹图谱构建及共有特征峰四级杆-静电场轨道阱高分辨质谱分析
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摘要
目的建立葛根药材的HPLC指纹图谱并对其共有特征峰进行高分辨质谱鉴定。方法葛根经30%(体积分数)乙醇提取,采用Inertsil ODS-3色谱柱(4.6 mm×250 mm,5μm),以乙腈-0.1%(体积分数)甲酸水溶液为流动相,梯度洗脱,检测波长为254 nm,对10批市售葛根药材的指纹图谱进行相似度评价分析,并通过对照品比对及四级杆-静电场轨道阱高分辨质谱(Q-Orbitrap HRMS/MS)对主要共有峰进行指认。结果建立了葛根的HPLC指纹图谱,10批葛根药材指纹图谱中有12个共有峰,各批次间共有峰的相对保留时间RSD均<0.5%,样品间相似度均>0.95,同时采用对照品比对法和高分辨液质技术对12个共有特征峰异黄酮成分进行了鉴定。结论所建立的葛根指纹图谱专属性强,结合采用高分辨液质联用技术进行的定性分析,可用于葛根药材的整体质量综合评价。
Objective To establish a quality assessment method for Puerariae Lobatae Radix based on HPLC fingerprint and qualitative analysis of the chemical compounds by HPLC-HRMS/MS. Methods The extract of Puerariae Lobatae Radix by 30%(volume fraction) ethanol was separated on an Inertsil ODS-3 column(250 mm × 4.6 mm,5 μm) with gradient elution. The mobile phase consisted of acetonitrile-0.1% formic acid. The detection wavelength was 254 nm. The similarity evaluation of 10 batches of Puerariae Lobatae Radix was carried out by the "Similarity Evaluation System for Chromatographic Fingerprints of TCM". HPLC-Q-Orbitrap HRMS/MS was used to characterize the chemical compounds of Puerariae Lobatae Radix. Results The chromatographic fingerprints of 10 batches of Puerariae Lobatae Radix were generated 12 common peaks. The relative retention time for 12 common peaks of each batch was less than 0.5%,and the similarity scores between each material batch and the reference fingerprint were greater than 0.95. A total of 12 components were identified by HPLC-Q-Orbitrap HRMS/MS. Conclusion The established method of fingerprint is specific and accurate,which can be used for quality evaluation of Puerariae Lobatae Radix.
Objective To establish a quality assessment method for Puerariae Lobatae Radix based on HPLC fingerprint and qualitative analysis of the chemical compounds by HPLC-HRMS/MS. Methods The extract of Puerariae Lobatae Radix by 30%(volume fraction) ethanol was separated on an Inertsil ODS-3 column(250 mm × 4.6 mm,5 μm) with gradient elution. The mobile phase consisted of acetonitrile-0.1% formic acid. The detection wavelength was 254 nm. The similarity evaluation of 10 batches of Puerariae Lobatae Radix was carried out by the "Similarity Evaluation System for Chromatographic Fingerprints of TCM". HPLC-Q-Orbitrap HRMS/MS was used to characterize the chemical compounds of Puerariae Lobatae Radix. Results The chromatographic fingerprints of 10 batches of Puerariae Lobatae Radix were generated 12 common peaks. The relative retention time for 12 common peaks of each batch was less than 0.5%,and the similarity scores between each material batch and the reference fingerprint were greater than 0.95. A total of 12 components were identified by HPLC-Q-Orbitrap HRMS/MS. Conclusion The established method of fingerprint is specific and accurate,which can be used for quality evaluation of Puerariae Lobatae Radix.
引文
[1] SONG W,LI Y,QIAO X,et al.Chemistry of the Chinese herbal medicine Puerariae Radix (Ge-Gen):a review[J].J Chin Pharm,2014,23(6):347-360.
[2] WONG K H,LI G Q,LI K M,et al.Kudzu root:traditional uses and potential medicinal benefits in diabetes and cardiovascular diseases[J].J Ethnopharmacol,2011,134(3):584-607.
[3] 张晓娟,周海纯.葛根化学成分,现代药理及临床应用研究进展[J].中医药信息,2017,34(1):128-130.
[4] 李克宁,郑惠婷,张雨,等.基于分子对接技术虚拟筛选葛根治疗缺血性脑卒中的物质基础研究[J].中草药,2018,49(8):1847-1853.
[5] 林卫东,胡靖敏,梁生旺,等.葛根改善胰岛素抵抗的网络药理学研究[J].中药材,2016,39(7):1628-1632.
[6] 林卫东,马文苑,田元新,等.基于分子对接技术探讨葛根改善胰岛素抵抗的物质基础[J].中国实验方剂学杂志,2016,22(6):194-199.
[7] 黄芳,祝婧云,梁新丽.葛根药材中化学成分HPLC指纹图谱研究[J].世界中医药,2016,11(12):2789-2792.
[8] 白而力.不同产地葛根HPLC指纹图谱研究[J].亚太传统医药,2015,11(13):24-26.
[9] 宋丽明,王文燕,张智超.陕西安康葛根药材的HPLC指纹图谱研究[J].中草药,2008,39(3):436-438.
[10] 李卓伦,王振辉,周霖,等.基于UPLC-Q-Orbitrap HRMS方法的丹黄祛瘀胶囊中多种化学成分研究[J].中草药,2017,48(18):3705-3713.
[11] 于舒婷,刘海霞,李昆,等.采用UHPLC-Q Exactive轨道阱高分辨质谱快速识别脑震宁颗粒的化学成分[J].药学学报,2018,53(4):609-620.
[12] 李静,李娟,贾金萍,等.基于UHPLC-Q Extractive轨道阱高分辨质谱的款冬花、叶的化学比较[J].药学学报,2018,53(3):444-452.
[13] 李晓明,杨滨,黄璐琦.高效液相色谱-质谱联用分析鉴别葛根的异黄酮成分[J].中国中药杂志,2008,33(11):1337-1339.
[14] 张启云,彭国梅,李冰涛,等.UHPLC-Q-TOF/MS技术分析葛根醇提液中化学成分[J].中药新药与临床药理,2017,28(4):513-518.
[15] DU G,ZHAO H Y,ZHANG Q W,et al.A rapid method for simultaneous determination of 14 phenolic compounds in Radix Puerariae using microwave-assisted extraction and ultra high performance liquid chromatography coupled with diode array detection and time-of-flight mass spectrometry[J].J Chromatogr A,2010,1217(5):705-714.
[2] WONG K H,LI G Q,LI K M,et al.Kudzu root:traditional uses and potential medicinal benefits in diabetes and cardiovascular diseases[J].J Ethnopharmacol,2011,134(3):584-607.
[3] 张晓娟,周海纯.葛根化学成分,现代药理及临床应用研究进展[J].中医药信息,2017,34(1):128-130.
[4] 李克宁,郑惠婷,张雨,等.基于分子对接技术虚拟筛选葛根治疗缺血性脑卒中的物质基础研究[J].中草药,2018,49(8):1847-1853.
[5] 林卫东,胡靖敏,梁生旺,等.葛根改善胰岛素抵抗的网络药理学研究[J].中药材,2016,39(7):1628-1632.
[6] 林卫东,马文苑,田元新,等.基于分子对接技术探讨葛根改善胰岛素抵抗的物质基础[J].中国实验方剂学杂志,2016,22(6):194-199.
[7] 黄芳,祝婧云,梁新丽.葛根药材中化学成分HPLC指纹图谱研究[J].世界中医药,2016,11(12):2789-2792.
[8] 白而力.不同产地葛根HPLC指纹图谱研究[J].亚太传统医药,2015,11(13):24-26.
[9] 宋丽明,王文燕,张智超.陕西安康葛根药材的HPLC指纹图谱研究[J].中草药,2008,39(3):436-438.
[10] 李卓伦,王振辉,周霖,等.基于UPLC-Q-Orbitrap HRMS方法的丹黄祛瘀胶囊中多种化学成分研究[J].中草药,2017,48(18):3705-3713.
[11] 于舒婷,刘海霞,李昆,等.采用UHPLC-Q Exactive轨道阱高分辨质谱快速识别脑震宁颗粒的化学成分[J].药学学报,2018,53(4):609-620.
[12] 李静,李娟,贾金萍,等.基于UHPLC-Q Extractive轨道阱高分辨质谱的款冬花、叶的化学比较[J].药学学报,2018,53(3):444-452.
[13] 李晓明,杨滨,黄璐琦.高效液相色谱-质谱联用分析鉴别葛根的异黄酮成分[J].中国中药杂志,2008,33(11):1337-1339.
[14] 张启云,彭国梅,李冰涛,等.UHPLC-Q-TOF/MS技术分析葛根醇提液中化学成分[J].中药新药与临床药理,2017,28(4):513-518.
[15] DU G,ZHAO H Y,ZHANG Q W,et al.A rapid method for simultaneous determination of 14 phenolic compounds in Radix Puerariae using microwave-assisted extraction and ultra high performance liquid chromatography coupled with diode array detection and time-of-flight mass spectrometry[J].J Chromatogr A,2010,1217(5):705-714.