不同加工麦冬中多元活性成分的含量测定及多元统计分析
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摘要
目的:建立超快速液相色谱-三重四极杆/线性离子阱质谱(UFLC-QTRAP-MS/MS)同时测定麦冬中甾体皂苷、高异黄酮、氨基酸及核苷类共32个成分含量的方法,考察不同加工方法对麦冬药材中多元活性成分的影响。方法:采用Synergi~(TM) Hydro-RP 100?柱(2.0 mm×100 mm,2.5μm),以0.1%甲酸水溶液-0.1%甲酸乙腈溶液为流动相,梯度洗脱,流速0.4 mL·min~(-1),柱温30℃,多反应离子监测(MRM)模式检测;用灰色关联度分析(GRA)和TOPSIS方法对多元活性成分进行综合评价。结果:32个成分在一定浓度范围内线性关系良好,相关系数均大于0.999 0;精密度、重复性、稳定性良好,RSD均小于5%;平均加样回收率为97.4%～102.0%,RSD均小于5%。GRA和TOPSIS分析结果显示,S5、S6样品综合质量较好。结论:所建立的方法准确、可靠,可用于麦冬药材内在质量的综合评价,该研究可为麦冬适宜产地加工方法的优选及其加工方法的规范化和标准化提供基础资料。
Objective:To investigate the influence of different processing methods on components in Ophiopogonis Radix,and establish a comprehensive analytical method using UFLC-QTRAP-MS/MS for the simultaneous determination of thirty-two components including steroidal saponins,homoisoflavonoids,amino acids and nucleosides in Ophiopogonis Radix. Methods:The chromatographic separation was performed on a SynergiTM Hydro-RP 100? column(2.0 mm×100 mm,2.5 μm) at 30 ℃ with a gradient elution of 0.1% formic acid solution-0.1%aqueous formic acid at a flow rate of 0.4 mL·min~(-1). The components were detected under multiple-reaction monitoring(MRM) mode. Besides,grey relation analysis(GRA) and TOPSIS methods have been applied to comprehensively analyze and evaluate the results of multiple active constituents. Results:The results showed that thirty-two components had good linearity with correlation coefficients above 0.999 0. The method exhibited good precision,repeatability and stability with the relative standard deviations(RSDs) less than 5%,the average recoveries were between 97.4% and 102.0% with RSDs below 5%. In addition,the results of GRA and TOPSIS indicated that the quality of S5 and S6 was better. Conclusion:The established method is accurate and reliable,which can be used to appraise the quality of Ophiopogonis Radix. Our study may lay the way for the processing method of Ophiopogonis Radix in optimization,normalization and standardization.
Objective:To investigate the influence of different processing methods on components in Ophiopogonis Radix,and establish a comprehensive analytical method using UFLC-QTRAP-MS/MS for the simultaneous determination of thirty-two components including steroidal saponins,homoisoflavonoids,amino acids and nucleosides in Ophiopogonis Radix. Methods:The chromatographic separation was performed on a SynergiTM Hydro-RP 100? column(2.0 mm×100 mm,2.5 μm) at 30 ℃ with a gradient elution of 0.1% formic acid solution-0.1%aqueous formic acid at a flow rate of 0.4 mL·min~(-1). The components were detected under multiple-reaction monitoring(MRM) mode. Besides,grey relation analysis(GRA) and TOPSIS methods have been applied to comprehensively analyze and evaluate the results of multiple active constituents. Results:The results showed that thirty-two components had good linearity with correlation coefficients above 0.999 0. The method exhibited good precision,repeatability and stability with the relative standard deviations(RSDs) less than 5%,the average recoveries were between 97.4% and 102.0% with RSDs below 5%. In addition,the results of GRA and TOPSIS indicated that the quality of S5 and S6 was better. Conclusion:The established method is accurate and reliable,which can be used to appraise the quality of Ophiopogonis Radix. Our study may lay the way for the processing method of Ophiopogonis Radix in optimization,normalization and standardization.
引文
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[14]贾诚,叶正良,姜秀晶,等.HPLC同时测定麦冬药材中3种黄酮类成分的含量[J].中国药学杂志,2011,46(15):1209JIA C,YE ZL,JIANG XJ,et al.Simultaneous determination of contents of three flavonoid ingredients in Radix Ophiopogonis by HPLC[J].Chin Pharm J,2011,46(15):1209
[15]吴发明,蔡晓洋,王盼,等.HPLC同时测定麦冬中5个皂苷成分的含量[J].中国中药杂志,2015,40(20):4022WU FM,CAI XY,WANG P,et al.HPLC simultaneous determination of contents of 5 saponin constituents in Ophiopogonis Radix[J].China J Chin Mater Med,2015,40(20):4022
[16]WANG Y,XU J,QU H,et al.Determination of three steroidal saponins from Ophiopogon japonicus(Liliaceae)via highperformance liquid chromatography with mass spectrometry[J].Nat Prod Res,2013,27(1):72
[17]WU Y,DONG Z,WU H,et al.Comparative studies on Ophiopogonis and Liriopes based on the determination of 11 bioactive components using LC-MS/MS and Hierarchical clustering analysis[J].Food Res Int,2014,57(1):15
[18]王胜男,华愉教,邹立思,等.不同加工玄参药材中多元功效成分的含量测定及灰色关联度分析[J].质谱学报,2017,38(3):328WANG SN,HUA YJ,ZOU LS,et al.Determination of multiple active constituents of Scrophulariae Radix with different processed products and grey relational analysis[J].J Chin Mass Spectrom Soc,2017,38(3):328
[19]王胜男,华愉教,邹立思,等.不同“发汗”条件加工玄参中核苷类成分的含量测定及灰色关联度分析[J].药物分析杂志,2016,36(6):1027WANG SN,HUA YJ,ZOU LS,et al.Content determination and grey relational analysis of nucleosides and nucleobases in different“sweating”processed products of Scrophulariae Radix[J].Chin JPharm Anal,2016,36(6):1027
[2]中华人民共和国药典2015年版.一部[S].2015:155ChP 2015.VolⅠ[S].2015:155
[3]LUN W,JIANG XL,ZHANG WM,et al.Homo-aro-cholestane,furostane and spirostane saponins from the tubers of Ophiopogon japonicas[J].Phytochemistry,2017,136:125
[4]CHEN MH,CHEN XJ,WANG M,et al.Ophiopogon japonicus-Aphytochemical,ethnomedicinal and pharmacological review[J].JEthnopharmacol,2016,181:193
[5]CHEN S,LI X,LIU L,et al.Ophiopogonin D alleviates high-fat diet-induced metabolic syndrome and changes the structure of gut microbiota in mice[J].Faseb J,2018,32(3):1139
[6]ISHIBASHI H,MOCHIDOME T,OKAI J,et al.Activation of potassium conductance by ophiopogonin-D in acutely dissociated rat paratracheal neurons[J].Br J Pharm,2001,132(2):461
[7]ZHAO JW,CHEN DS,DENG CS,et al.Evaluation of antiinflammatory activity of compounds isolated from the rhizome of Ophiopogon japonicas[J].BMC Complement Altern Med,2017,17(1):7
[8]HE F,XU BL,CHEN C,et al.Methylophiopogonanone Asuppresses ischemia/reperfusion-induced myocardial apoptosis in mice via activating PI3K/Akt/eNOS signaling pathway[J].Acta Pharmacol Sin,2016,37(6):763
[9]WANG Y,LIU F,LIANG Z,et al.Homoisoflavonoids and the antioxidant activity of ophiopogon japonicus root[J].Iran J Pharm Res,2017,16(1):357
[10]张黄琴,刘培,朱邵晴,等.双边栝楼成熟果实不同部位游离氨基酸及核苷类资源性化学成分分析与评价[J].中国现代中药,2017,19(12):1683ZHANG HQ,LIU P,ZHU SQ,et al.Analysis and evaluation of amino acids and nucleosides in different parts of ripe fruit of Trichosanthes rosthornii Harm[J].Mod Chin Med,2017,19(12):1683
[11]段金廒,宿树兰,吕洁丽,等.药材传统加工传统经验与现代科学认识[J].中国中药杂志,2009,34(24):3151DUAN JA,SU SL,L JL,et al.Traditional experiences and modern cognition on primary processing of traditional Chinese medicinal materials[J].China J Chin Mater Med,2009,34(24):3151
[12]吴发明,张芳芳,李敏,等.川麦冬产地干燥方法综合评价研究[J].中药材,2015,38(7):1400WU FM,ZHANG FF,LI M,et al.Study on comprehensive evaluation of drying methods of sichuan Ophiopogon japonicus at production area[J].J Chin Med Mater,2015,38(7):1400
[13]吴发明,曾俊,李敏,等.基于药材性状特征和含量测定分析干燥温度对麦冬质量的影响[J].药物分析杂志,2016,36(7):1310WU FM,ZENG J,LI M,et al.Effect of drying temperature on quality of Ophiopogonis Radix based on the appearance character characteristics and content of chemical components[J].Chin JPharm Anal,2016,36(7):1310
[14]贾诚,叶正良,姜秀晶,等.HPLC同时测定麦冬药材中3种黄酮类成分的含量[J].中国药学杂志,2011,46(15):1209JIA C,YE ZL,JIANG XJ,et al.Simultaneous determination of contents of three flavonoid ingredients in Radix Ophiopogonis by HPLC[J].Chin Pharm J,2011,46(15):1209
[15]吴发明,蔡晓洋,王盼,等.HPLC同时测定麦冬中5个皂苷成分的含量[J].中国中药杂志,2015,40(20):4022WU FM,CAI XY,WANG P,et al.HPLC simultaneous determination of contents of 5 saponin constituents in Ophiopogonis Radix[J].China J Chin Mater Med,2015,40(20):4022
[16]WANG Y,XU J,QU H,et al.Determination of three steroidal saponins from Ophiopogon japonicus(Liliaceae)via highperformance liquid chromatography with mass spectrometry[J].Nat Prod Res,2013,27(1):72
[17]WU Y,DONG Z,WU H,et al.Comparative studies on Ophiopogonis and Liriopes based on the determination of 11 bioactive components using LC-MS/MS and Hierarchical clustering analysis[J].Food Res Int,2014,57(1):15
[18]王胜男,华愉教,邹立思,等.不同加工玄参药材中多元功效成分的含量测定及灰色关联度分析[J].质谱学报,2017,38(3):328WANG SN,HUA YJ,ZOU LS,et al.Determination of multiple active constituents of Scrophulariae Radix with different processed products and grey relational analysis[J].J Chin Mass Spectrom Soc,2017,38(3):328
[19]王胜男,华愉教,邹立思,等.不同“发汗”条件加工玄参中核苷类成分的含量测定及灰色关联度分析[J].药物分析杂志,2016,36(6):1027WANG SN,HUA YJ,ZOU LS,et al.Content determination and grey relational analysis of nucleosides and nucleobases in different“sweating”processed products of Scrophulariae Radix[J].Chin JPharm Anal,2016,36(6):1027