Herein, a strategy is proposed for the simultaneous determination of primary coumarins in Peucedani Radix(Chinese name: Qianhu). The methodology consists of three consecutive steps: 1) Semi-preparative LC in combinati...Herein, a strategy is proposed for the simultaneous determination of primary coumarins in Peucedani Radix(Chinese name: Qianhu). The methodology consists of three consecutive steps: 1) Semi-preparative LC in combination with a home-made automated fraction collection module to fragment the universal metabolome standard into ten fractions(Frs. I–X); 2) LC–accurate MS/MS and quantitative1 H NMR spectroscopy conducted in parallel to acquire the qualitative and quantitative data of each fraction; 3) Robust identification and quantification of components by use of LC coupled to multiple reaction monitoring. In this final step, the most significant fractions(Frs. III–X) were pooled to serve as the pseudo-mixed standard solution. Meticulous online parameter optimization was performed to obtain the optimal parameters, including ion transitions and collision energies. Concerns were particularly paid onto pursuing the parameters being capable of monitoring regiospecific isomers, notably praeruptorin E vs. 3′-isovaleryl-4′-angeloylkhellactone. The quantitative performance of the method was validated according to diverse assays. Eleven primary coumarins(1–11) were unambiguously identified and absolutely quantified, even though no external reference compound was used. Above all, the integrated strategy not only provides a feasible pipeline for the quality assessment of Peucedani Radix, but more importantly, shows the potential for authentic compound-free quantitative evaluation of traditional Chinese medicines.展开更多
为探究白花前胡和紫花前胡中代谢物的种类及相对含量,借助于广泛靶向代谢组学的方法,利用UPLC-MS技术对白花前胡和紫花前胡成分进行分析、鉴定,确定主要差异代谢产物,并对其进行相对定量分析;从整体角度出发,结合多元统计分析,对二者中...为探究白花前胡和紫花前胡中代谢物的种类及相对含量,借助于广泛靶向代谢组学的方法,利用UPLC-MS技术对白花前胡和紫花前胡成分进行分析、鉴定,确定主要差异代谢产物,并对其进行相对定量分析;从整体角度出发,结合多元统计分析,对二者中主要差异性成分进行系统鉴定,并对差异代谢物进行KEGG通路富集分析。从白花前胡和紫花前胡中共鉴定出12类1556个代谢产物,主要差异代谢物12类共1085种,其中酚酸类、黄酮类、木脂素及香豆素类、生物碱类和脂类等化合物差异较大,分别占比为19.2%、15.9%、10.1%、9.4%和9.2%;其中421种成分上调,664种成分下调。差异代谢产物以biosynthesis of phenylpropanoids(ko01061)、linoleic acidmetabolism(ko00591)、pentose phosphate pathway(ko00030)、biosynthesis of secondarymetabolites(ko01110)、phosphotransferase system(ko02060)、phenylalanine和Tyrosine and tryptophan biosynthesis(ko00400)等6条通路富集显著,主要为酚酸类、脂肪酸类等成分的关键代谢通路。研究表明,白花前胡和紫花前胡化学成分差异较大,2020版《中国药典》将其单列为不同的药材,具有一定的先进性。本研究可为白花前胡和紫花前胡的标准修订及品质评价提供参考。展开更多
为应用ITS2序列对前胡、紫花前胡药材及其混伪品进行DNA条形码鉴定研究,该文依据国家药典委员会公布的《中药材DNA条形码分子鉴定指导原则》,PCR扩增前胡、紫花前胡及其混伪品的ITS2序列并进行双向测序。经CodonCode Aligner V 3.7.1对...为应用ITS2序列对前胡、紫花前胡药材及其混伪品进行DNA条形码鉴定研究,该文依据国家药典委员会公布的《中药材DNA条形码分子鉴定指导原则》,PCR扩增前胡、紫花前胡及其混伪品的ITS2序列并进行双向测序。经CodonCode Aligner V 3.7.1对测序峰图进行校对拼接,采用MEGA5.0软件分析相关数据。结果表明：前胡药材的ITS2序列长度为229~230 bp,种内变异较小,平均K2P遗传距离为0.005。紫花前胡药材的ITS2序列长度为227 bp,种内无变异。前胡、紫花前胡药材与其混伪品的种间平均K2P遗传距离分别为0.044,0.065。紫花前胡的种间最小K2P遗传距离明显大于其种内的最大K2P遗传距离。最小距离法和NJ树鉴定结果表明前胡、紫花前胡均能与其混伪品明显区分。因此,ITS2序列可有效鉴别前胡、紫花前胡药材及其混伪品。展开更多
基金financially supported by National Science Fund of China (Nos. 81773875,81403073 and 81530097)Quality Guarantee System of Chinese Herbal Medicines (No. 201507002)+1 种基金foundation from Beijing University of Chinese Medicine (No. 2016-JYB-XJQ004)the Macao Science and Technology Development Fund (007/2014/AMJ)
文摘Herein, a strategy is proposed for the simultaneous determination of primary coumarins in Peucedani Radix(Chinese name: Qianhu). The methodology consists of three consecutive steps: 1) Semi-preparative LC in combination with a home-made automated fraction collection module to fragment the universal metabolome standard into ten fractions(Frs. I–X); 2) LC–accurate MS/MS and quantitative1 H NMR spectroscopy conducted in parallel to acquire the qualitative and quantitative data of each fraction; 3) Robust identification and quantification of components by use of LC coupled to multiple reaction monitoring. In this final step, the most significant fractions(Frs. III–X) were pooled to serve as the pseudo-mixed standard solution. Meticulous online parameter optimization was performed to obtain the optimal parameters, including ion transitions and collision energies. Concerns were particularly paid onto pursuing the parameters being capable of monitoring regiospecific isomers, notably praeruptorin E vs. 3′-isovaleryl-4′-angeloylkhellactone. The quantitative performance of the method was validated according to diverse assays. Eleven primary coumarins(1–11) were unambiguously identified and absolutely quantified, even though no external reference compound was used. Above all, the integrated strategy not only provides a feasible pipeline for the quality assessment of Peucedani Radix, but more importantly, shows the potential for authentic compound-free quantitative evaluation of traditional Chinese medicines.
文摘为探究白花前胡和紫花前胡中代谢物的种类及相对含量,借助于广泛靶向代谢组学的方法,利用UPLC-MS技术对白花前胡和紫花前胡成分进行分析、鉴定,确定主要差异代谢产物,并对其进行相对定量分析;从整体角度出发,结合多元统计分析,对二者中主要差异性成分进行系统鉴定,并对差异代谢物进行KEGG通路富集分析。从白花前胡和紫花前胡中共鉴定出12类1556个代谢产物,主要差异代谢物12类共1085种,其中酚酸类、黄酮类、木脂素及香豆素类、生物碱类和脂类等化合物差异较大,分别占比为19.2%、15.9%、10.1%、9.4%和9.2%;其中421种成分上调,664种成分下调。差异代谢产物以biosynthesis of phenylpropanoids(ko01061)、linoleic acidmetabolism(ko00591)、pentose phosphate pathway(ko00030)、biosynthesis of secondarymetabolites(ko01110)、phosphotransferase system(ko02060)、phenylalanine和Tyrosine and tryptophan biosynthesis(ko00400)等6条通路富集显著,主要为酚酸类、脂肪酸类等成分的关键代谢通路。研究表明,白花前胡和紫花前胡化学成分差异较大,2020版《中国药典》将其单列为不同的药材,具有一定的先进性。本研究可为白花前胡和紫花前胡的标准修订及品质评价提供参考。
