基于指纹图谱及多指标成分定量分析的肿节风配方颗粒质量标准研究
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摘要
目的建立肿节风配方颗粒(SHDG)HPLC指纹图谱及多指标成分定量分析方法,比较不同厂家及批次SHDG的质量差异,为SHDG的质量控制提供依据。方法采用HPLC-UV法,以乙腈-0.2%甲酸水溶液为流动相,梯度洗脱,建立SHDG指纹图谱,并利用对照品和HPLC-Q/TOF法对其特征峰进行指认;同时建立绿原酸、异嗪皮啶、迷迭香酸的含量测定方法。采用Chempattern化学计量学软件对结果进行处理分析。通过聚类分析和主成分分析,将不同厂家及相同厂家不同批次的制剂进行分类,并阐明造成差异的主要成分。结果建立SHDG指纹图谱,确证了7个特征峰并进行了指认,分别是新绿原酸、绿原酸、隐绿原酸、咖啡酸、异嗪皮啶、迷迭香酸-4-O-β-D-葡萄糖苷、迷迭香酸。55 min内SHDG的主要色谱峰能够达到完全分离;绿原酸、异嗪皮啶、迷迭香酸分别在各自范围内峰面积与质量浓度呈良好的线性关系;平均回收率分别为98.92%(RSD 1.54%)、98.20%(RSD 1.12%)、99.58%(RSD 1.12%);18批样品绿原酸质量分数为0.33～1.39 mg/g,异嗪皮啶质量分数为1.31～2.74 mg/g,迷迭香酸质量分数为1.11～4.54 mg/g;18批样品与共有模式的相似度为0.688～0.992;不同厂家及批次的SHDG中绿原酸、异嗪皮啶、迷迭香酸含量存在较大的差异。结论根据所建立的指纹图谱,结合3个主要成分含量测定能够为SHDG的质量控制提供更全面的参考。
Objective To establish HPLC and multiple components determination method of Sarcandrae Herba Dispensing Granules(SHDG), in order to compare the difference of the quality in various SHDG samples and provide an effective method to ensure the quality of SHDG. Methods HPLC-UV method was used to establish the characteristic chromatogram of SHDG, and acetonitrile-0.2% formic acid solution was used as the mobile phase with the gradient elution. The common peaks were identified by comparison with the reference standards and HPLC-Q/TOF. At the same time, the method for simultaneous determination of chlorogenic acid, isofraxidin, and rosmarinic acid was established with the same approach. Chemometrics software Chempattern was employed to analyze the data. Through cluster analysis and principal component analysis, different preparations from different manufacturers and different batches from the same manufacturer were classified, and the main components causing the differences were clarified. Results The SHDG fingerprint was established to confirm and identify seven characteristic peaks, namely,neochlorgenic acid, chlorogenic acid, cryptochlorogenic acid, caffeic acid, isofraxidin, rosmarinic acid-4-O-β-D-glucoside, and rosmarinic acid. The main chromatographic peaks of SHDG can be completely separated within 55 min. There was a good linear relationship between the peak area and concentration of chlorogenic acid, isofraxidin, and rosmarinic acid. The average recovery rates were 98.92%(RSD 1.54%), 98.20%(RSD 1.12%), and 99.58%(RSD 1.12%), respectively. The chlorogenic acid content of 18 batches of samples was 0.33—1.39 mg/g, the content of isocyanidine was 1.31—2.74 mg/g, and the content of rosmarinic acid was1.11—4.54 mg/g. The similarity between 18 batches of samples and the common mode was 0.688—0.992. There were certain differences in the content of chlorogenic acid, isofraxidin, rosmarinic acid in the SHDG of various batches and manufacturers.Conclusion The proposed specific HPLC characteristic chromatogram and quantitation method of three components for SHDG offered more comprehensive reference for quality control of the crude drug.
Objective To establish HPLC and multiple components determination method of Sarcandrae Herba Dispensing Granules(SHDG), in order to compare the difference of the quality in various SHDG samples and provide an effective method to ensure the quality of SHDG. Methods HPLC-UV method was used to establish the characteristic chromatogram of SHDG, and acetonitrile-0.2% formic acid solution was used as the mobile phase with the gradient elution. The common peaks were identified by comparison with the reference standards and HPLC-Q/TOF. At the same time, the method for simultaneous determination of chlorogenic acid, isofraxidin, and rosmarinic acid was established with the same approach. Chemometrics software Chempattern was employed to analyze the data. Through cluster analysis and principal component analysis, different preparations from different manufacturers and different batches from the same manufacturer were classified, and the main components causing the differences were clarified. Results The SHDG fingerprint was established to confirm and identify seven characteristic peaks, namely,neochlorgenic acid, chlorogenic acid, cryptochlorogenic acid, caffeic acid, isofraxidin, rosmarinic acid-4-O-β-D-glucoside, and rosmarinic acid. The main chromatographic peaks of SHDG can be completely separated within 55 min. There was a good linear relationship between the peak area and concentration of chlorogenic acid, isofraxidin, and rosmarinic acid. The average recovery rates were 98.92%(RSD 1.54%), 98.20%(RSD 1.12%), and 99.58%(RSD 1.12%), respectively. The chlorogenic acid content of 18 batches of samples was 0.33—1.39 mg/g, the content of isocyanidine was 1.31—2.74 mg/g, and the content of rosmarinic acid was1.11—4.54 mg/g. The similarity between 18 batches of samples and the common mode was 0.688—0.992. There were certain differences in the content of chlorogenic acid, isofraxidin, rosmarinic acid in the SHDG of various batches and manufacturers.Conclusion The proposed specific HPLC characteristic chromatogram and quantitation method of three components for SHDG offered more comprehensive reference for quality control of the crude drug.
引文
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[13]郭烨,吴安明,何杨虎,等.不同产地不同采收期肿节风药材主要活性成分含量比较[J].江西中医药,2019(1):65-67.
[14]宋晓虹,李小艳,李成仁,等.肿节风药材HPLC指纹图谱研究[J].天然药物研究与开发,2009,21(1):111-113.
[15]姚志红,王其意,韦卓纯,等.超高效液相色谱法同时测定肿节风中6个成分的含量[J].分析测试学报,2016,35(6):641-647.
[16]罗永东,林培玲,吴水生,等.应用ASE-LC-ESI-MS法快速筛选肿节风的化学成分[J].亚太传统医药,2011,7(12):29-31.
[17]Yan D H,Liu X D,Deng X M,et al.Quality assessment of crude and processed Leonuri Fructus by chemical and color analysis combined with chemometric method[J].Chin Herb Med,2018,10(4):388-395.
[18]周霖,周珮珮,王肖辉,等.UPLC-Q-Orbitrap HRMS结合主成分分析的丹灯通脑软胶囊质量评价研究[J].中草药,2018,49(18):4311-4318.
[19]Shen J,Li P,He C N,et al.Simultaneous determination of 15 flavonoids from different parts of Scutellaria baicalensis and its chemometrics analysis[J].Chin Herb Med,2019,11(1):20-27.
[2]王刚力,陈道峰,林瑞超.肿节风的化学成分及其制剂质量控制研究进展[J].中草药,2003,34(8):12-14.
[3]王冰岚,童胜强,颜继忠.肿节风注射液研究进展及临床应用[J].中成药,2007,29(3):426-429.
[4]邹小燕,高慧媛,吴斌,等.肿节风化学成分的研究[J].中草药,2007,38(3):354-356.
[5]宋朝阳.肿节风注射液联合阿帕替尼治疗晚期胃癌的临床研究[J].现代药物与临床,2017,32(6):1114-1117.
[6]易洁梅.肿节风的药理研究及临床应用新进展[J].实用药物与临床,2011,14(6):523-525.
[7]王力福,沈美勤,赵金花.肿节风分散片联合洛美沙星治疗急性化脓性中耳炎的疗效观察[J].现代药物与临床,2016,31(8):1260-1263.
[8]Swarup V,Ghosh J,Ghosh S,et al.Antiviral and neuroprotective role of Octaguanidinium dendrimerconjugated morpholino oligomers in Japanese encephalitis[J].Antimicrob Agents Chemother,2007,51(9):3367-3370.
[9]Prasad N R,Jeyanthimala K,Ramachandran S.Coffeic acid modulates ultraviolet radiation-B induced oxidative damage in human blood lymphocytes[J].J Photochem Photobiol B,2009,95(3):196-203.
[10]程会,汤庆发,陈飞龙,等.UPLC测定不同粒度肿节风有效成分的含量[J].中药新药与临床药理,2014,25(4):480-483.
[11]邓伟麟,吴莹莹,毕丹,等.肿节风药材的HPLC指纹图谱研究及其异嗪皮啶与迷迭香酸的含量测定[J].中国药房,2013,24(47):4472-4474.
[12]姚志红,潘宇明,戴毅,等.肿节风水提物及其市售制剂的HPLC指纹图谱研究[J].中草药,2010,41(9):1457-1460.
[13]郭烨,吴安明,何杨虎,等.不同产地不同采收期肿节风药材主要活性成分含量比较[J].江西中医药,2019(1):65-67.
[14]宋晓虹,李小艳,李成仁,等.肿节风药材HPLC指纹图谱研究[J].天然药物研究与开发,2009,21(1):111-113.
[15]姚志红,王其意,韦卓纯,等.超高效液相色谱法同时测定肿节风中6个成分的含量[J].分析测试学报,2016,35(6):641-647.
[16]罗永东,林培玲,吴水生,等.应用ASE-LC-ESI-MS法快速筛选肿节风的化学成分[J].亚太传统医药,2011,7(12):29-31.
[17]Yan D H,Liu X D,Deng X M,et al.Quality assessment of crude and processed Leonuri Fructus by chemical and color analysis combined with chemometric method[J].Chin Herb Med,2018,10(4):388-395.
[18]周霖,周珮珮,王肖辉,等.UPLC-Q-Orbitrap HRMS结合主成分分析的丹灯通脑软胶囊质量评价研究[J].中草药,2018,49(18):4311-4318.
[19]Shen J,Li P,He C N,et al.Simultaneous determination of 15 flavonoids from different parts of Scutellaria baicalensis and its chemometrics analysis[J].Chin Herb Med,2019,11(1):20-27.