HPLC指纹图谱、Q-TOF/MS定性及多成分定量相结合的大黄饮片质量评价研究
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摘要
目的建立更全面的大黄饮片质量评价方法。方法 HPLC法建立35批大黄饮片指纹图谱,并生成对照指纹图谱,标定共有峰,并进行相似度评价;Q-TOF/MS对共有峰进行鉴定,对通过对照品比对确认的13个蒽醌类成分进行定量测定。结果大黄饮片指纹图谱标定共有峰45个,包括蒽醌类成分17个(其中芦荟大黄素-8-O-葡萄糖苷、大黄酸-8-O-葡萄糖苷、大黄素-1-O-葡萄糖苷、大黄酚-1-O-葡萄糖苷、大黄酚-8-O-葡萄糖苷、芦荟大黄素-3-羟甲基-O-葡萄糖苷、大黄素-8-O-葡萄糖苷、大黄素甲醚-8-O-葡萄糖苷8个结合型蒽醌及芦荟大黄素、大黄酸、大黄素、大黄酚、大黄素甲醚5个游离型蒽醌通过对照品比对确认)、蒽酮类成分2个(番泻苷B和番泻苷A,均通过对照品比对确认)、鞣质类成分17个(其中没食子酸、儿茶素、表没食子儿茶素没食子酸酯、表儿茶素、表儿茶素没食子酸酯通过对照品比对确认)、二苯乙烯类成分2个[白藜芦醇4′-O-葡萄糖苷和白藜芦醇4′-O-β-D-(6″-O-没食子酰)-葡萄糖苷,均通过对照品比对确认]、苯丁酮类成分4个[其中4′-羟基苯基-2-丁酮-4′-O-β-D-(2″-O-没食子酰-6″-O-对羟基桂皮酰)-葡萄糖苷通过对照品比对确认]、色原酮类成分2个、萘类成分1个;35批大黄饮片中13个蒽醌类成分含量测定结果差异较大。结论建立的方法可用于大黄饮片质量评价。
Objective To establish a more comprehensive quality evaluation method for rhubarb pieces. Methods Fingerprints of 35 batches of rhubarb pieces were determined by HPLC, the control fingerprint was established, the common peaks were calibrated and the similarity was evaluated. Components of common peaks were identified by Q-TOF/MS and the content of 13 anthraquinones confirmed by reference substance was determined. Results There were 45 common peaks in the fingerprint of rhubarb pieces,including 17 anthraquinones(among them, eight bound anthraquinones of aloe-emodin-8-O-glucoside, rhein-8-O-glucoside, emodin-1-O-glucoside, chrysophanol-1-O-glucoside, chrysophanol-8-O-glucoside, aloe-emodin-3-hydroxymethyl-O-glucoside, emodin-8-Oglucoside, physcion-8-O-glucoside and five free anthraquinones of aloe-emodin, rhein, emodin, chrysophanol, physcion were confirmed by reference substance), two anthrones(sennoside B and sennoside A, confirmed by reference substances), 17 tannins(among them, gallic acid, catechin, epigallocatechin gallate, epicatechin and epicatechin gallate were confirmed by reference substance), two stilbenes [resveratrol 4′-O-glucoside and resveratrol 4′-O-β-D-(6″-O-galloyl)-glucoside, confirmed by reference substances], four butyrophenone [among them, 4-(4′-hydroxyphenyl)-2-butanone 4′-O-β-D-(2″-O-galloyl-6″-O-p-hydroxy-cinnamyl)glucoside, confirmed by reference substances], two chromones and one naphthalenes. The content of 13 anthraquinones in 35 batches of rhubarb pieces was quite different. Conclusion The established method can be used to evaluate the quality of rhubarb pieces.
Objective To establish a more comprehensive quality evaluation method for rhubarb pieces. Methods Fingerprints of 35 batches of rhubarb pieces were determined by HPLC, the control fingerprint was established, the common peaks were calibrated and the similarity was evaluated. Components of common peaks were identified by Q-TOF/MS and the content of 13 anthraquinones confirmed by reference substance was determined. Results There were 45 common peaks in the fingerprint of rhubarb pieces,including 17 anthraquinones(among them, eight bound anthraquinones of aloe-emodin-8-O-glucoside, rhein-8-O-glucoside, emodin-1-O-glucoside, chrysophanol-1-O-glucoside, chrysophanol-8-O-glucoside, aloe-emodin-3-hydroxymethyl-O-glucoside, emodin-8-Oglucoside, physcion-8-O-glucoside and five free anthraquinones of aloe-emodin, rhein, emodin, chrysophanol, physcion were confirmed by reference substance), two anthrones(sennoside B and sennoside A, confirmed by reference substances), 17 tannins(among them, gallic acid, catechin, epigallocatechin gallate, epicatechin and epicatechin gallate were confirmed by reference substance), two stilbenes [resveratrol 4′-O-glucoside and resveratrol 4′-O-β-D-(6″-O-galloyl)-glucoside, confirmed by reference substances], four butyrophenone [among them, 4-(4′-hydroxyphenyl)-2-butanone 4′-O-β-D-(2″-O-galloyl-6″-O-p-hydroxy-cinnamyl)glucoside, confirmed by reference substances], two chromones and one naphthalenes. The content of 13 anthraquinones in 35 batches of rhubarb pieces was quite different. Conclusion The established method can be used to evaluate the quality of rhubarb pieces.
引文
[1]谭鹏,张海珠,张青,等.UPLC法同时测定大黄中10个蒽醌衍生物的含量[J].中草药,2018,49(4):928-934.
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[5]陈林伟,秦昆明,徐雪松,等.中药指纹图谱数据库的研究现状及展望[J].中草药,2014,45(21):3041-3047.
[6]Chen F F,Qi H Y,Shi Y P.Fingerprint analysis of Codonopsis Radix by HPLC coupled with chemometrics analysis[J].Chin Herb Med,2013,5(4):307-312.
[7]秦建平,吴建雄,郎悦,等.痛宁凝胶的HPLC-MS指纹图谱研究[J].现代药物与临床,2016,31(4):427-430.
[8]李莉,李德坤,鞠爱春.注射用五味子提取物指纹图谱研究[J].药物评价研究,2018,41(3):475-480.
[9]王月红,游飞祥,王磊,等.六经头痛片HPLC指纹图谱建立及多指标成分定量测定研究[J].中草药,2017,48(20):4167-4173.
[10]窦志华,乔进,卞理,等.指纹图谱与一测多评法相结合的大黄质量控制方法[J].中国药学杂志,2015,50(5):442-448.
[11]游飞祥,袁雪海,许浚,等.基于HPLC-Q-TOF/MS的六经头痛片化学成分分析[J].中草药,2017,48(20):4157-4166.
[12]窦志华,卞理,许波,等.指纹图谱、多成分定量与模式识别相结合的大黄质量评价[J].中华中医药学刊,2017,35(11):2903-2907.
[13]洪筱坤,王智华,李旭,等.19个大黄样品HPLC指纹图谱的比较分析[J].上海中医药杂志,1993,27(8):32-34.
[14]Lin C C,Wu C I,Lin T C,et al.Determination of 19rhubarb constituents by high performance liquid chromatography-ultraviolet-mass spectrometry[J].J Sep Sci,2006,29(17):2584-2593.
[15]Jin W,Wang Y F,Ge R L,et al.Simultaneous analysis of multiple bioactive constituents in Rheum tanguticum Maxim.ex Balf.by high-performance liquid chromatography coupled to tandem mass spectrometry[J].Rapid Commun Mass Specttrom,2007,21(14):2351-2360.
[16]Ye M,Han J,Chen H B,et al.Analysis of phenolic compounds in rhubarbs using liquid chromatography coupled with electrospray ionization mass spectrometry[J].J Am Soc Mass Spectrom,2007,18(1):82-91.
[17]Zhu T T,Liu X,Wang X L,et al.Profiling and analysis of multiple compounds in rhubarb decoction after processing by wine steaming using UHPLCQ-TOF-MS coupled with multiple statistical strategies[J].J Sep Sci,2016,39(15):3081-3090.
[18]Yan Y,Zhang Q,Feng F.HPLC-TOF-MS and HPLC-MS/MS combined with multivariate analysis for the characterization and discrimination of phenolic profiles in nonfumigated and sulfur-fumigated rhubarb[J].J Sep Sci,2016,39(14):2667-2677.
[19]王晴,卢志威,刘月红,等.UPLC-Q-TOF/MSE结合诊断离子过滤方法快速分析大黄中酚类成分[J].中国中药杂志,2017,42(10):1922-1931.
[20]Zhang L,Liu H,Qin L,et al.Global chemical profiling based quality evaluation approach of rhubarb using ultra performance liquid chromatography with tandem quadrupole time-of-flight mass spectrometry[J].J Sep Sci,2015,38(3):511-522.
[21]Liu Y,Li L,Xiao Y Q,et al.Global metabolite profiling and diagnostic ion filtering strategy by LC-QTOF MS for rapid identification of raw and processed pieces of Rheum palmatum L.[J].Food Chem,2016,192:531-540.
[22]Wang M,Fu J F,Guo H M,et al.Discrimination of crude and processed rhubarb products using a chemometric approach based on ultra fast liquid chromatography with ion trap/time-of-flight mass spectrometry[J].J Sep Sci,2015,38(3):395-401.
[23]郑俊华,西泽信,山岸乔,等.正品大黄中34种化学成分的高效液相色谱法定量分析[J].北京医科大学学报,1989,21(1):54-56.
[24]Kashiwada Y,Nonaka G I,Nishioka I.Studies on rhubarb(Rhei Rhizoma).XV1).Simultaneous determination of phenolic constituents by high-performance liquid chromatography[J].Chem Pharm Bull,1989,37(4):999-1004.
[25]Komatsu K,Nagayama Y,Tanaka K,et al.Comparative study of chemical constituents of rhubarb from different origins[J].Chem Pharm Bull,2006,54(11):1491-1499.
[26]赵倩,陈育鹏,崔旭盛,等.掌叶大黄UPLC多指标成分测定及指纹图谱研究[J].药物分析杂志,2018,38(10):1697-1710.
[27]梁淋渊,甘麦邻,罗燕.大黄化学成分与药理活性研究进展[J].中兽医医药杂志,2017,36(1):80-83.
[28]Michodjehoun-Mestres L,Amraoui W,Brillouet J M.Isolation,characterization,and determination of 1-O-trans-cinnamoyl-β-D-glucopyranose in the epidermis and flesh of developing cashew apple(Anacardium occidentale L.)and four of its genotypes[J].J Agric Food Chem,2009,57(4):1377-1382.
[29]刘月红,黄政海,董玲,等.高效液相色谱法同时测定大黄中14种成分的含量[J].中国中药杂志,2017,42(23):4514-4519.
[30]Wang Z W,Wang J S,Luo J,et al.Three new phenolic glucosides from the roots of Rheum palmatum[J].Chem Pharm Bull,2012,60(2):241-245.
[31]窦志华,曹瑞,卞理,等.正交试验法优选大黄中蒽醌类成分提取工艺[J].中草药,2018,49(14):3279-3286.
[32]Chen A Z,Sun L,Yuan H,et al.Simultaneous qualitative and quantitative analysis of 11 active compounds in rhubarb using two reference substances by UHPLC[J].JSep Sci,2018,41(19):3686-3696.
[33]李丽.大黄炮制前后物质基础变化规律研究[D].北京:中国中医科学院,2011.
[34]刘翠玲,刘东辉,黄月纯,等.大黄饮片HPLC指纹图谱的方法学研究[J].中药新药与临床药理,2009,20(5):442-445.
[35]陶明宝,张乐,刘飞,等.含蒽醌类成分中药的安全性研究进展[J].中药药理与临床,2016,32(6):238-243.
[2]龚小红,赵梦杰,党珏,等.基于主成分分析不同产地大黄13个成分量的比较研究[J].中草药,2017,48(23):4994-4999.
[3]李冬华,张平,张明童,等.全国大黄药材及其饮片评价性抽验结果分析与一测多评法研究[J].中国药事,2018,32(6):775-783.
[4]任伟光,王琦,黄林芳,等.大黄类药材的质量评价进展[J].中南药学,2014,12(4):354-359.
[5]陈林伟,秦昆明,徐雪松,等.中药指纹图谱数据库的研究现状及展望[J].中草药,2014,45(21):3041-3047.
[6]Chen F F,Qi H Y,Shi Y P.Fingerprint analysis of Codonopsis Radix by HPLC coupled with chemometrics analysis[J].Chin Herb Med,2013,5(4):307-312.
[7]秦建平,吴建雄,郎悦,等.痛宁凝胶的HPLC-MS指纹图谱研究[J].现代药物与临床,2016,31(4):427-430.
[8]李莉,李德坤,鞠爱春.注射用五味子提取物指纹图谱研究[J].药物评价研究,2018,41(3):475-480.
[9]王月红,游飞祥,王磊,等.六经头痛片HPLC指纹图谱建立及多指标成分定量测定研究[J].中草药,2017,48(20):4167-4173.
[10]窦志华,乔进,卞理,等.指纹图谱与一测多评法相结合的大黄质量控制方法[J].中国药学杂志,2015,50(5):442-448.
[11]游飞祥,袁雪海,许浚,等.基于HPLC-Q-TOF/MS的六经头痛片化学成分分析[J].中草药,2017,48(20):4157-4166.
[12]窦志华,卞理,许波,等.指纹图谱、多成分定量与模式识别相结合的大黄质量评价[J].中华中医药学刊,2017,35(11):2903-2907.
[13]洪筱坤,王智华,李旭,等.19个大黄样品HPLC指纹图谱的比较分析[J].上海中医药杂志,1993,27(8):32-34.
[14]Lin C C,Wu C I,Lin T C,et al.Determination of 19rhubarb constituents by high performance liquid chromatography-ultraviolet-mass spectrometry[J].J Sep Sci,2006,29(17):2584-2593.
[15]Jin W,Wang Y F,Ge R L,et al.Simultaneous analysis of multiple bioactive constituents in Rheum tanguticum Maxim.ex Balf.by high-performance liquid chromatography coupled to tandem mass spectrometry[J].Rapid Commun Mass Specttrom,2007,21(14):2351-2360.
[16]Ye M,Han J,Chen H B,et al.Analysis of phenolic compounds in rhubarbs using liquid chromatography coupled with electrospray ionization mass spectrometry[J].J Am Soc Mass Spectrom,2007,18(1):82-91.
[17]Zhu T T,Liu X,Wang X L,et al.Profiling and analysis of multiple compounds in rhubarb decoction after processing by wine steaming using UHPLCQ-TOF-MS coupled with multiple statistical strategies[J].J Sep Sci,2016,39(15):3081-3090.
[18]Yan Y,Zhang Q,Feng F.HPLC-TOF-MS and HPLC-MS/MS combined with multivariate analysis for the characterization and discrimination of phenolic profiles in nonfumigated and sulfur-fumigated rhubarb[J].J Sep Sci,2016,39(14):2667-2677.
[19]王晴,卢志威,刘月红,等.UPLC-Q-TOF/MSE结合诊断离子过滤方法快速分析大黄中酚类成分[J].中国中药杂志,2017,42(10):1922-1931.
[20]Zhang L,Liu H,Qin L,et al.Global chemical profiling based quality evaluation approach of rhubarb using ultra performance liquid chromatography with tandem quadrupole time-of-flight mass spectrometry[J].J Sep Sci,2015,38(3):511-522.
[21]Liu Y,Li L,Xiao Y Q,et al.Global metabolite profiling and diagnostic ion filtering strategy by LC-QTOF MS for rapid identification of raw and processed pieces of Rheum palmatum L.[J].Food Chem,2016,192:531-540.
[22]Wang M,Fu J F,Guo H M,et al.Discrimination of crude and processed rhubarb products using a chemometric approach based on ultra fast liquid chromatography with ion trap/time-of-flight mass spectrometry[J].J Sep Sci,2015,38(3):395-401.
[23]郑俊华,西泽信,山岸乔,等.正品大黄中34种化学成分的高效液相色谱法定量分析[J].北京医科大学学报,1989,21(1):54-56.
[24]Kashiwada Y,Nonaka G I,Nishioka I.Studies on rhubarb(Rhei Rhizoma).XV1).Simultaneous determination of phenolic constituents by high-performance liquid chromatography[J].Chem Pharm Bull,1989,37(4):999-1004.
[25]Komatsu K,Nagayama Y,Tanaka K,et al.Comparative study of chemical constituents of rhubarb from different origins[J].Chem Pharm Bull,2006,54(11):1491-1499.
[26]赵倩,陈育鹏,崔旭盛,等.掌叶大黄UPLC多指标成分测定及指纹图谱研究[J].药物分析杂志,2018,38(10):1697-1710.
[27]梁淋渊,甘麦邻,罗燕.大黄化学成分与药理活性研究进展[J].中兽医医药杂志,2017,36(1):80-83.
[28]Michodjehoun-Mestres L,Amraoui W,Brillouet J M.Isolation,characterization,and determination of 1-O-trans-cinnamoyl-β-D-glucopyranose in the epidermis and flesh of developing cashew apple(Anacardium occidentale L.)and four of its genotypes[J].J Agric Food Chem,2009,57(4):1377-1382.
[29]刘月红,黄政海,董玲,等.高效液相色谱法同时测定大黄中14种成分的含量[J].中国中药杂志,2017,42(23):4514-4519.
[30]Wang Z W,Wang J S,Luo J,et al.Three new phenolic glucosides from the roots of Rheum palmatum[J].Chem Pharm Bull,2012,60(2):241-245.
[31]窦志华,曹瑞,卞理,等.正交试验法优选大黄中蒽醌类成分提取工艺[J].中草药,2018,49(14):3279-3286.
[32]Chen A Z,Sun L,Yuan H,et al.Simultaneous qualitative and quantitative analysis of 11 active compounds in rhubarb using two reference substances by UHPLC[J].JSep Sci,2018,41(19):3686-3696.
[33]李丽.大黄炮制前后物质基础变化规律研究[D].北京:中国中医科学院,2011.
[34]刘翠玲,刘东辉,黄月纯,等.大黄饮片HPLC指纹图谱的方法学研究[J].中药新药与临床药理,2009,20(5):442-445.
[35]陶明宝,张乐,刘飞,等.含蒽醌类成分中药的安全性研究进展[J].中药药理与临床,2016,32(6):238-243.